Difference between revisions of "Team:Freiburg/NotebookDelivery"

Latest revision as of 23:18, 19 October 2016

Lab Journals

Group 5 - Enzymatic activity and Sporeshampoo

The focus of this group lays on the confirmation of the enzymatic activity of our spores. To show that the GST is correctly expressed by the spores a GST-assay is adjusted to the use in a plate reader. Another application of the spores would be for a (anti-dandruff) shampoo, so this is tested by washing a GFP-coated surface with different detergents and our construct and measuring the fluorescence.

05.08.2016

plate coated with GFP:

Prepared coating solution: used protocol from “Abeam: direct Elisa coating”. Took 0.78 g Na2CO3, 1.5 g NaHCO3 and dissolved it in 250 ml H2O, pH: 9.5.
Dilute stock solution of GFP (1350µg/ml) in coating buffer for coating concentration of 20 µg/ml.
Coated 93 wells with 50µl coating solution
Incubated for 2 h at room temperature
Washed the wells twice with PBS (V=200 µl)
Blocked it with 5% nonfat dry milk in PBS (2.5 g in 50 ml PBS), blocked coated wells with 200 µl solution.
Incubated for 2 h at room temperature.
Washed blocked wells twice with PBS (V=200 µl)

Measuring:

Measured the emission from top, Measured the emission from bottom

06.08.2016

Measured the plate from 05.08.2016 again with only one blank:

fig.1: The Average over replicates of fluorescence in RFU. Measured the fluorescence of coated GFP (20 µg/ml) and the auto fluorescence of empty wells as a negative control, by measuring from top with the plate reader.

fig.2: The Average over replicates of fluorescence in RFU. Measured the fluorescence of coated GFP (20 µg/ml) and the auto fluorescence of empty wells as a negative control, by measuring from bottom with the Plate reader.

Prepared coating solutions with several concentrations:

20 µg/ml
28.7 µg/ml
41.8 µg/ml
35.6 µg/ml

07.08.2016

Coated MED. Binding Greiner Plate:

Coated the wells with 50 µl from prepared coating solution:

fig.3: The Average over replicates of fluorescence in RFU. Measured the fluorescence of coated GFP (20 µg/ml, 28.7 µg/ml, 41.8 µg/ml and 35.6 µg/ml) and the auto fluorescence of empty wells as a negative control, by measuring from top with the Plate reader.

fig.4: The Average over replicates of fluorescence in RFU. Measured the fluorescence of coated GFP (20 µg/ml, 28.7 µg/ml, 41.8 µg/ml and 35.6 µg/ml), the auto fluorescence of empty wells as a negative control and the fluorescence of GFP (20 µg/ml, 28.7 µg/ml, 41.8 µg/ml and 35.6 µg/ml) in solution as positive control, by measuring from top with the plate reader.

08.08.2016

Coated

Used Greiner MID bind plate
Spores: WT, B53, B54; used 50 µl/well, spores were purified by washing with PBS, to remove the medium.
used GFP coating concentrations of: 40 µg/ml, 45 µg/ml, 60 µg/ml, 70 µg/ml, 80 µg/ml, 90 µg/ml; used 50 µl/well
m.Cherry: 20 µg/ml, 40 µg/ml, 80 µg/ml

coated over night at 4°C in the fridge.

09.08.2016

Blocked

Washed plate from previous day twice with 200 µl PBS
Incubated for 2 h at room temperature
Blocked with 200 µl 5% nonfat dry milk in PBS
Incubated for 2 h at room temperature

Measured

Measured the fluorescence at two different focal heights:
22mm:

fig.5: Average of fluorescence over replicates of samples (B53_coated 18 mill., B53_solution 18 mill., B53_coated 9 mill., B53_solution 9 mill., B53_coated 1.8 mill., B53_solution 1.8 mill., B54_cated 18 mill., B54_solution 18 mill., B54_coated 9 mill., B54_solution 9 mill., B54_coated 1.8 mill., B54_solution 1.8 mill., GFP_coated 20 µg/ml, GFP_coated 35.6 µg/ml, GFP_coated 40 µg/ml, GFP_coated 45 µg/ml, GFP_coated 60 µg/ml, GFP_coated 70 µg/ml, GFP_coated 80 µg/ml, GFP_coated 90 µg/ml, m.Cherry_coated 20 µg/ml, m.Cherry_coated 40 µg/ml, m.Cherry_coated 80 µg/ml, WT_coated 18 mill.) and an empty well as negative control, at a focal height of 22 mm.

2 mm:

fig.6: Average of fluorescence over replicates of samples (B53_coated 18 mill., B53_solution 18 mill., B53_coated 9 mill., B53_solution 9 mill., B53_coated 1.8 mill., B53_solution 1.8 mill., B54_cated 18 mill., B54_solution 18 mill., B54_coated 9 mill., B54_solution 9 mill., B54_coated 1.8 mill., B54_solution 1.8 mill., GFP_coated 20 µg/ml, GFP_coated 35.6 µg/ml, GFP_coated 40 µg/ml, GFP_coated 45 µg/ml, GFP_coated 60 µg/ml, GFP_coated 70 µg/ml, GFP_coated 80 µg/ml, GFP_coated 90 µg/ml, m.Cherry_coated 20 µg/ml, m.Cherry_coated 40 µg/ml, m.Cherry_coated 80 µg/ml, WT_coated 18 mill.) and an empty well as negative control, at a focal height of 2 mm.

13.08.2016

Dried in PBS

Scheme:

fig.7: Pipetting scheme for the measuring of fluorescence at the surface with several concentrations of GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml) and two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54). The shaded wells show wells with PBS dried, where the solution of GFP/spores was dried at 37°C in the incubator.

took from B54 and B53 aliquot 108 µl (108 mill. Spores) and 54µl (54 mill. spores)
washed them by spinning down at 13000rpm for 13min and resuspended the pellet in PBS, repeated three times
resuspended final pellet in 300µl PBS -pipetted 50 µl solution into the shaded wells (scheme)
incubated at 37°C for 5,5 h

Measured

Measured the plate with different focal heights and gain values:

Focal height 22 mm, gain 1740
Focal height self-adjusted at 3.6 mm, gain 1740
Focal height 3.6 mm, gain 2960

Result

Dry

fig.8: Average of relative fluorescence over replicates of dried in PBS, GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml), two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54) and empty wells as negative control. Used setting: focal height of 22 mm and gain value of 1740.

fig.9: Average of relative fluorescence over replicates of dried in PBS, GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml), two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54) and empty wells as negative control. Used setting: focal height of 3.6 mm and gain value of 1740.

fig.10: Average of relative fluorescence over replicates of dried in PBS, GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml), two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54) and empty wells as negative control. Used setting: focal height of 3.6 mm and gain value of 2960.

Wet:

fig.11: Average of relative fluorescence over replicates with PBS on top GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml), two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54) and empty wells as negative control. It was used a setting with a focal height of 22 mm and gain value of 1740.

fig.12: Average of relative fluorescence over replicates with PBS on top GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml), two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54) and empty wells as negative control. It was used a setting with a focal height of 3.6 mm and gain value of 1740.

fig.13: Average of relative fluorescence over replicates with PBS on top GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml), two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54) and empty wells as negative control. Used setting: focal height of 3.6 mm and gain value of 2960.

14.08.2016 – 15.08.2016

Dried in H2O

scheme:

fig.14: Pipette scheme for the measuring of fluorescence at the surface by several concentrations of GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml) and two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54). The shaded wells shown dried wells with H2O, were the solution of GFP/spores was dried by 37°C in the incubator.

Preparation:

Prepared dilution series of GFP and spores in H2O as in the scheme
Pipetted 100 µl per well and incubated it at 37°C for 12 h 40 min.

Measured:

Measured the plate with three different settings:
* Focal height 22 mm, gain 1740 * Focal height 3.6 mm, gain 1740 * Focal height 3.6 mm, gain 2960

Result:

Focal height 22 mm, gain 1740:

fig.15: Average of relative fluorescence over replicates of dried in H2O, GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml), two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54) and empty wells as negative control. Used setting: focal height of 22 mm and gain value of 1740.

Focal height 3.6 mm, gain 1740:

fig.16: Average of relative fluorescence over replicates of dried in H2O, GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml), two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54) and empty wells as negative control. It was used a setting with a focal height of 3.6 mm and gain value of 1740.

Focal height 3.6 mm, gain 2960:

fig.17: Average of relative fluorescence over replicates of dried in H2O, GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml), two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54) and empty wells as negative control. Used setting: focal height of 3.6 mm and gain value of 2960.

16.08.2016 – 15. 08.2016

Coated

Coated free wells on the plate where GFP dried in with PBS, used wells D5 – D12, H5 – H12 and D1 as blank
Coated concentrations of: 20 µg/ml (D5 – D7), 15 µg/ml (D8 – D10), 10 µg/ml (D11, D12 and H5), 5 µg/ml (H6 – H8)
Coated with 50 µl
Incubated for 2 h at room temperature
Washed wells twice with 200 µl PBS
Blocked with 200 µl 5% nonfat dry milk in PBS
Incubated for 2 h at room temperature
Washed the wells twice with 200 µl PBS

Measured

Measured the emission of the fluorescence with several settings:
Focal height 22 mm, gain 1740
Focal height 3.6 mm, gain 174
Focal height 3.6 mm, gain 2960

Result

Focal height 22 mm, gain 1740:

fig.18: Measured the relative fluorescence over replicates of coated GFP (20 µg/ml, 15 µg/ml, 10 µg/ml, 5 µg/ml) and empty wells as negative control, at focal height 22 mm with a gain value of 1740.

Focal height 3.6 mm, gain 1740:

fig.19: Measured the relative fluorescence over replicates of coated GFP (20 µg/ml, 15 µg/ml, 10 µg/ml, 5 µg/ml) and empty wells as negative control, at focal height 3.6 mm with a gain value of 1740.

fig.20: Measured the relative fluorescence over replicates of coated GFP (20 µg/ml, 15 µg/ml, 10 µg/ml, 5 µg/ml) and empty wells as negative control, at focal height 3.6 mm with a gain value of 2960.

19.08.2016

Plate coated and washed

Coated

Used Greiner HIGH bind plate
Coating concentration of GFP 20 µg/ml
Coating volume 50 µl
Incubated for 4 h at room temperature

Blocked

Washed coated wells twice with 200 µl PBS
Blocked with 200 µl 5% nonfat dry milk in PBS
Incubated for 2 h at room temperature
Washed wells twice with 200 µl PBS

Measured

Measured with a focal height of 3.6 mm and a gain value of 1740

Result

fig.20: Average of relative fluorescence units over replicates of coated GFP (20 µg/ml) and empty wells as negative control.

Washing – Assay 01

scheme:

fig.21: washed coated GFP with different detergents (SDS, Tx100, Tween20, Shampoo1 (Syoss OLEO 21) and Shampoo 2 (Herbal Essences Verwöhnende Feuchtigkeit), one group was washed with 2% SDS as positive control, empty was washed with PBS and wasn’t coated with GFP, the positive control was washed with PBS.

Concentration for each detergent:
SDS: 2%; 0,1%; 0,01%; 0,001%; 0,0001%
Triton: 0,1%; 0,01%; 0,001%; 0,0001%
Tween: 0,1%; 0,01%; 0,001%; 0,0001%
Shampoo 1: 0,1%; 0,01%; 0,001%; 0,0001%
Shampoo 2: 0,1%; 0,01%; 0,001%; 0,0001%

Washed the plate after the scheme with 200 µl detergent and shook for 2 min, pipetted out.
Measured the fluorescence of GFP
Washed with PBS
Measured the fluorescence of GFP
Washed again with 200 µl detergent and shook for 2 min, pipetted out
Measured the fluorescence of GFP
Washed with PBS
Measured the fluorescence of GFP

Result

21.08.2016

Washing – Assay 02

Prepared different concentrations of detergents:

SDS: 2%; 1,5%; 1%; 0,5%; 0,1%
Triton: 1,5%; 1%; 0,5%; 0,1%
Tween: 1,5%; 1%; 0,5%; 0,1%
Shampoo 1: 1,5%; 1%; 0,5%; 0,1%
Shampoo 2: 1,5%; 1%; 0,5%; 0,1%

For washing used plate and scheme from 19.06.2016 with different concentrations.
washed the coated wells with the detergents, pipetted 200 µl detergent and shook for 2 min, pipetted out.
Measured the fluorescence emission after washing with detergent.
Measured the fluorescence emission after washing with PBS.

Measured

after washing with detergents
washed with PBS afterwards

Results

fig.24: RFU difference between last measurement of the plate (washed again with PBS) and washed with detergents: SDS (2%; 1,5%; 1%; 0,5%; 0,1%), Triton: 1,5%; 1%; 0,5%; 0,1%), Tween (1,5%; 1%; 0,5%; 0,1%), Shampoo 1 (1,5%; 1%; 0,5%; 0,1%), Shampoo 2 (1,5%; 1%; 0,5%; 0,1%), SDS 2% was used as positive control. Used a focal height of 3.6 mm and a gain value of 1740.

23.08.2016

GST-Assay 01

Preparation

Prepared dilution of GST stock (5 Units/ml):

1 U/ml
0.75 U/ml
0.5 U/ml
0.1 U/ml
0.02 U/ml
0.001 U/ml

Volume master mix 1.5 ml: (15 µlCDNB [100 mM] + 15 µl GSH [100 mM] + 1470 µl PBS).

Plate scheme:

fig.25: scheme for measuring the activity of different GST concentrations (1 U/ml, 0.75 U/ml, 0.5 U/ml, 0.1 U/ml, 0.02 U/ml, 0.001 U/ml), Empty (PBS and master mix), blank (PBS and master mix).

Measured

start the reaction with 30 µl GST as in the scheme (Abb.27), in blank and empty (control) 30 µl PBS was added.
Setrtings:protocol_gst_run_1_.xlsx

Result

All data and graphs were collected in an Excel file: freiburg_2016_gst_run_1.xlsx

GST-Assay 02

Preparation

prepared samples:

0.3 U (2x 0.15 U) 0.15 U (90 µl of 5 U/ml stock)
0.25 U (2x 0.125 U) 0.125 U (25 µl of 5 U/ml stock + 5 µl PBS)
0.2 U (2x 0.1 U) 0.1 U (20 µl of 5 U/ml stock + 10 µl PBS)
0.15 U (2x 0.075 U) 0.075 U (15 µl of 5 U/ml stock + 15 µl PBS)
0.1 U (2x 0.05 U) 0.05 U (10 µl of 5 U/ml stock + 20 µl PBS)
0.05 U (2x 0.025 U) 0.025 U (5 µl of 5 U/ml stock + 25 µl PBS)
0.02 U (2x 0.01 U) 0.01 U (2 µl of 5 U/ml stock + 28 µl PBS)

volume of each sample contains 30 µl.
Prepared mester mix (1470 µl PBS + 15 µl GSH [100 mM] + 15 µl CDNB [100 mM]).
Pipetted GST samples like in the scheme (fig.26)

Scheme:

fig.26: pipetting scheme of GST amount (0.15 U, 0.125 U, 0.1 U, 0.075 U, 0.05 U, 0.025 U, 0.01 U), white=empty (master mix and PBS) and black=blank (master mix and PBS).

Measured

Started the reaction with 90 µl master mix in each well
Put it immediately in the plate reader
Used measurement settings:protocol_gst_run_2_.xlsx

Results

All data and graphs were collected in an Excel file:freiburg_2016_18.09.16_gst-assay_02_.xlsx

GST – Assay 03

Repeated the assay of GST – assay 01
Pipetting scheme as in GST – assay 01 (fig.26)

Measured

Start the reaction with 90 µl master mix in each well
Used settings as shown in the reader protocol: protocol_gst_run_3.xlsx

Results

All graphs were collected in an Excel file: freiburg_2016_gst_run3.xlsx

GST – Assay 04

Repeat the assay depending on the results of GST – assay 03 (Freiburg 2016_gst_run3.xlsx). increased the measuring time up to 10 min and pipetted on ice.
Pipetted after scheme shown in fig.26.

Measured

Start the reaction with 90 µl master mix in each well
Put it immediately in the plate reader
Used settings as shown in the reader protocol: protocol_gst_run_4.xlsx

Results

All graphs were collected in an Excel file: freiburg_2016_gst_run4.xlsx

29.08.2016

Detergent screening

Scheme:

fig.27: scheme of detergent screening for SDS, Tx100, Tween20, Shampoo1 and Shampoo2, as positive control a solution of 2% SDS was used, as negative control a wash solution of PBS without detergents was used.

Coated

Prepared GFP coating solution (20 µg/ml)
Coated the plate after the scheme (fig.27) with coating volume of 100 µl
Incubated for 2 h at room temperature
Washed the wells twice with 200 µl PBS
Blocked wells with 200 µl 5% nonfat dry milk in PBS
Incubated for 2 h at room temperature
Washed wells twice with 200 µl PBS

30.08.2016

Preparation of detergents for screening: Prepared stock solutions of 20% and 40%
- Prepared samples of 1 ml for each of the three cycles in each block (except block three there were only two cycles):

block 1: 2, 4, 6, 8% solutions of Tx100, Tween20, Shampoo1 (SH1) and Shampoo2 (SH2); for SDS used concentrations 1.5, 0.25, 1, 0.5 and 0.1% solutions
block 2: 12, 14, 16,1 8% solutions of Tx100, Tween20, Shampoo1 (SH1) and Shampoo2 (SH2); for SDS used concentrations 1.5, 0.75, 1, 0.5 and 0.1% solutions
block 3: 32, 34,3 6, 38% solutions of Tx100, Tween20, Shampoo1 (SH1) and Shampoo2 (SH2); for SDS used concentrations 1.5, 1.25, 1, 0.5 and 0.1% solutions

Washing

Block 1:

Washed with solutions with concentrations of 2, 4, 6, 8% of Tx100, Tween20, Shampoo1 (SH1) and Sampoo2 (SH2); for SDS used concentrations 1.5, 0.25, 1, 0.5 and 0.1%, for negative control used PBS and for positive control 2% SDS. Wash solution volume took 200 µl.
washed with PBS twice afterwards
pipetted out the whole volume which was left in the wells
measured the fluorescence
repeated it for a total of 3 cycles

block 2:

Washed with solutions with concentrations of 12,14, 16, 18% of Tx100, Tween20, Shampoo1 (SH1) and Shampoo2 (SH2); for SDS used concentrations 1.5, 0.75, 1, 0.5 and 0.1%, for negative control used PBS and for positive control 2% SDS. Wash solution volume took 200 µl.
washed with PBS twice afterwards
pipetted out the whole volume which was left in the wells
measured the fluorescence
repeated it for a total of 3 cycles

block 3:

Washed with solutions with concentrations of 32,34, 36, 38% of Tx100, Tween20, Shampoo1 (SH1) and Shampoo2 (SH2); for SDS used concentrations 1.5, 1.25, 1, 0.5 and 0.1%, for negative control used PBS and for positive control 2% SDS. Wash solution volume took 200 µl.
washed with PBS twice afterwards
pipetted out the whole volume which was left in the wells
measured the fluorescence
repeated it

Result

Data and graphs were collected in an Excel file:freiburg_2016_30.08.16_detergengt_screening.xlsx

Fig.28: wash efficiency of detergents in an overview.

02.09.2016

Washing – Assay 03

Plate name W5
coated the plate with GFP after coating protocol, used scheme from fig.29

scheme:

Fig.29: washing scheme with several concentrations of detergents: SDS: 1.25, 1, 0.75, 0.25%; Tween20: 22, 24, 26, 28, 32, 24, 26, 28%; SH1: 12, 14, 16 ,18, 22, 24, 26, 28%. For neg. contr. used 2% SDS, pos. contr. was washed with PBS without detergent.

stock solution 40%

measured

used setting for measuring the fluorescence from coated GFP and washed plate as shown in the protocol:

Result

Detailed figures collected in an Excel file:

Fig.30: overview of the wash efficiency of detergents: SDS: 1.25, 1, 0.75, 0.25%; Tween20: 22, 24, 26, 28, 32, 24, 26, 28%; SH1: 12, 14, 16 ,18, 22, 24, 26, 28%. For neg. contr. used 2% SDS, pos. contr. was washed with PBS without detergent.

GST – Assay 05

repeated the assay depending on the results of GST – assay 04 (Excel file: Freiburg 2016gst_run4.xlsx.

scheme:

fig.26: pipetting scheme of GST amounts (0.15 U, 0.125 U, 0.1 U, 0.075 U, 0.05 U, 0.025 U, 0.01 U), empty (master mix and PBS); blank (master mix and PBS).

prepared the samples on ice
incubated the plate before starting in the fridge for 5 min.
prepared samples after the scheme shown in fig.26

Measured

started the reaction with 90 µl master mix in each well
put it immediately in the plate reader
used settings as shown in the reader protocol: protocol_gst_run_5.xlsx

Result

All data and graphs were collected in an excel file: freiburg_2016_gst_run5.xlsx

03.09.2016

GST – assay 06

Preparation as in GST – assay 05. Took a little more time to start the measuring after mixing enzyme and master mix
Used pipetting scheme as shown in fig.26.

Measuring

Used setting as shown in the reader protocol: protocol_gst_run_6.xlsx

Result

All data and graphs were collected in an Excel file: freiburg_2016_gst_run6.xlsx

GST – Assay 07

Repeated like as in assay 05, but everything was pipetted on ice. Used pipetting scheme as shown in fig.26.

Measured

Start the reaction with 90 µl master mix from low GST concentration to high, for reducing the time delay the master mix was pipetted directly next to the plate reader

Result

All data and graphs were collected in an Excel file: freibur_2016_gst_run7.xlsx

GST – Assay 08

Preparation

Preparations like in the other assays before (GST – assay 07) after pipetting and incubating for 5 min in the fridge, put the plate on a shaker.

Measuring

started the reaction with 90 µl master mix while shaking at 300rpm and shaker temperature of 12°C.
Put it then into the reader.
protocol: protocol_gst_run_8.xlsx

Result

graphs were collected in an Excel file: freiburg_2016_gst_run8.xlsx

GST-Assay 09

Preparation

* as in GST-assay 8 * increased measuring time up to 20 min in the measuring settings

Measuring

Started the reaction like in GST-assay 8, used settings as shown in the protocol: protocol_gst_run_9.xlsx

Result

All data and graphs were in an Excel file: freiburg_2016_gst_run9.xlsx

12.09.2016

GST – Assay 10

Preparation

samples of GST:

0.15 U (90 µl of 5 U/ml stock)
0.075 U (15µl of 5 U/ml stock + 75 µl PBS)
0.025 U (5 µl of 5 U/ml stock + 85 µl PBS)
0,005 U (3 µl of 5 U/ml stock + 87 µl PBS)

master mix: 15 µl CDNB + 15 µl GSH + 1470 µl PBS.
Used 30 µl enzyme solution and 90 µl master mix per well, for blank and empty used 30 µl PBS and master mix.

Scheme:

fig.31: Pipetting scheme for GST – assay with 0.15, 0.075, 0.025 and 0.005 Units of GST and master mix, for wells empty and blank used instead of GST solutions PBS and master mix.

master mix wasn’t enough for all wells, concentration of 0.15 U without master mix.

Measured

Used setting as shown in reader protocol:

Result

Data and graphs were collected in an Excel file:

GST – assay 11

Repeated GST – assay 10 with more master mix, scheme fig.31
1,7 ml: 17 µl CDNB + 17 µl GSH + 1666 µl PBS

Measured

Used setting as shown in reader protocol:

Result

Data and graphs were collected in an Excel file:

GST – assay 12

Repeated measurement of GST – assay 11 with different measurement settings.
Used the same scheme fig.31

Measured

Used setting as shown in reader protocol:

Result

Data and graphs were collected in an Excel file:

14.09.2016

Shampoo: washing with nanobody

Prepared

Coated plate after the scheme
Prepared nanobody solution: Concentration nanobody stock solution 0.48 mg/ml, set up dilution of 1.8 ng/µl
Scheme:

fig.32: washing scheme for washing with nanobody and detergents. As control were used: GFP washed with PBS (negative control 1), washed with nanobody in PBS (negative control 2), 2%SDS (positive control). As reference empty wells were not coated and unwashed, same as the blank. Used detergent samples: SDS: 1, 0.75, 0.5, 0.25% solutions with nanobody, Tween20: 22, 24, 26, 28% solutions with nanobody.

wash – assay

Measured the fluorescence of coated GFP
Washed with 1 µl nanobody and 200 µl concentrations of detergents: SDS: 1.5, 1, 0.5, 0.25%, Tween20: 22, 24, 26, 28%, SH1: 12, 14, 16, 18%
Incubated it for 5 min
Shook it for 5 min at 300 rpm
washed with PBS twice afterwards
pipetted out the whole volume which was left in the wells
measured the fluorescence, used same settings as measured the fluorescence of coated GFP:

Result

Detailed column charts were collected in an Excel file: washing_with_nanobody_eff..xlsx

fig.33: overview of the efficiency of washing with nanobody in combination with detergents. SDS: 1, 0.75, 0.5, 0.25% solutions with nanobody, Tween20: 22, 24, 26,28% solutions with nanobody. Controls GFP washed with PBS (negative control 1), washed with nanobody in PBS (negative control 2), 2%SDS (positive control).

14.09.2016

GST – Assay 13

Scheme:

fig.34: pipetting scheme of GST – assay, used GST amount of: 0.0001, 0.00001, 0.000001, 0.0000001 Units, for empty and blank was used PBS instead of GST.

Measured

Measurement settings as in the reader protocol:14.09.16_gst-assay_settings.xlsx

Result

All data and graphs were collected in an excel file: freiburg_2016_14.09.16_gst-assay_-1.xlsx

16.09.2016

Shampoo

Platename: WN2
Scheme:
Fig.35: washing scheme for washing with nanobody and detergents. As control were used: coated GFP not washed (negative control 1), GFP washed with PBS (negative control 2), washed with nanobody in PBS (negative control 3), 2%SDS (positive control). As reference were empty wells not coated and unwashed, same as the blank. Used detergent samples: SDS: 1, 0.75, 0.5, 0.25% solutions with nanobody, Tween20: 22, 24, 26,28% solutions with nanobody.

Prepared

coated plate with GFP (coating concentration 20 µg/ml)
dilute 1 µl nanobody stock solution (0.48 mg/ml) in prepared samples 1 ml detergents: SDS: 1, 0.75, 0.5, 0.25, Tween20: 22, 24, 26, 28%, SH1: 12, 14, 16, 18%

Wash-assay

Measured the fluorescence of coated GFP
Washed with 200 µl detergents: SDS: 1.5, 1, 0.5, 0.25% (row A and B with nanobody, row C and D without nanobody), Tween20: 22, 24, 26, 28% (row A and B with nanobody, row C and D without nanobody), SH1: 12, 14, 16, 18% (row E and F with nanobody, row G and H without nanobody)
Incubated it for 5 min
Shook it for 5 min at 300 rpm
washed with PBS twice afterwards
pipetted out the whole volume which was left in the wells
measured the fluorescence
used same settings as measured the fluorescence of coated GFP:

Result

Detailed column charts were collected in an Excel file:freiburg_2016_16.09.16_washing_nanobody_02_gfp_master.xlsx

Fig.36: Overview of the wash efficiency with and without nanobody for: SDS, SDS + nanobody, Tween20, Twenn20 + nanobody, SH1, SH1 + nanobody.

17.09.2016

GST – Assay 14

Repeat the GST – assay 13 with the same scheme (fig34).
Concentration of samples:

0,000 1U (3 µl 0,1 U/ml stock + 87 µl PBS)
0,00001 U (3 µl 0,01 U/ml stock + 87µl PBS)
0,000001 U (3 µl 0,001 U/ml stock + 87µl PBS)
0,0000001 U (3 µl 0,0001 U/ml stock + 87µl PBS)

master mix: 15 µl CDNB + 15 µl GSH + 1470 µl PBS.
Pipetted the GST after the scheme (30 µl per well), used for empty wells and blank instead of GST solution PBS.

Measured

Start the reaction with 90 µl mastr mix from low concentration to high concentration, pipetted next to the plate reader to reduce the time delay.
Measured the absorbance with settings as described in the measurement protocol: 17.09.16_gst-assay_01_settings.xlsx

GST – assay 15

Repeated the GST – assay 14
settings: 17.09.16_gst-assay_02_settings.xlsx

GST – assay 16

Changed setting for the measurement: shook in plate reader before each cycle at 500 rpm 1 s.

Scheme:

fig.37: GST – assay pipetting scheme for GST concentrations of 0.005 and 0.0000001 Units. Empty as negative control with PBS instead of GST, blank with the same conditions like empty.

prepared

samples of 0.005 and 0.0000001 U
master mix (14 µl CDNB + 14 µl GSH + 1372 µl PBS)
pipetted GST samples after the scheme and PBS for the controls (30 µl)

Measurement

start the reaction with 170 µl master mix. Used new setting as in the reader protocol described: 17.09.16_gst-assay_03_settings.xlsx

Result

All data and graphs of GST - assay 14 - 16 were collected in an Excel file: freiburg_2016_17.09.16_gst-assay_master.xlsx

18.09.2016

GST – Assay 17

Doubled the amount of substrate
Scheme:

Fig.38: Pipetting scheme for GST – assay with 0.1, 0.025, 0.005 and 0.001 Units (U), empty wells and blank measured with PBS instead of GST as control.

Prepared

GST samples:

0.1 U (60 µl of 5 U/ml stock + 30 µl PBS)
0.025 U (5 µl of 5 U/ml stock + 87 µl PBS)
0.005 U (3 µl of 5 U/ml stock + 85 µl PBS)
0,001 U (3 µl of 1 U/ml stock + 87 µl PBS)

master mix: 30 µl CDNB + 30 µl GSH + 1440 µl PBS.
Pipet GST samples and PBS after the scheme (fig.38).

Measured

Start the reaction with 90 µl master mix
Start measuring with settings which are described in the protocol: 18.09.16_gst-assay_01_settings.xlsx

Result

Data and graphs were collected in an Excel file:18.09.16_gst-assay_01.xlsx

GST – assay 18

Scheme:

fig.39: Pipetting scheme for GST assay with 0.1, 0.05 Units (U), empty wells and blank measured with PBS instead of GST as control, grey wells were not used.

Increased the measuring time up to 33 minutes.

Prepared

GST Samples:

0,1 U
0,05 U (30 µl of 5 U/ml stock + 60 µl PBS)
0,025 U

master mix

Measurement

Start reactions with 90 µl master mix. Used setting as described in the measurement protocol of the reader: 18.09.16_gst-assay_02_settings.xlsx

Result

Data and graphs were collected in an Excel file: freiburg_2016_18.09.16_gst-assay_02_-1.xlsx

19.09.2016

GST – assay 19

Scheme:

fig.40: Pipetting scheme for GST – assay with 0.1, 0.05 Units (U), empty wells and blank measured with PBS instead of GST as control, grey wells were not used.

Prepared

GST concentrations:

0,1 U (80 µl 5 U/ml stock + 40 µl PBS)
0,05 U (40 µl 5 U/ml stock + 80 µl PBS)

master mix: 18 µl CDNB + 18 µl GSH + 1164 µl PBS
Changed shaking time before the first cycle in the setting to 200 rpm for 1s
Pipetted the samples after the scheme in fig.42

Measurement

Start the reactions with 90 µl master mix and start measurement with the setting as described in the plate reader protocol: 19.09.16_gst-assay_01_settings.xlsx

GST – assay 20

Repeat the GST – assay 19 in addition with one well of enzyme without master mix
Scheme:

Fig.41: Pipetting scheme for GST – assay with 0.1, 0.05 Units (U), empty wells and blank measured with PBS instead of GST as control, En = well which contains only enzyme without assay cocktail, grey wells were not used.

Prepared

GST concentrations:

0,1 U (80 µl 5 U/ml stock + 40 µl PBS)
0,05 U (40 µl 5 U/ml stock + 80 µl PBS)

master mix: 18 µl CDNB + 18 µl GSH + 1164 µl PBS
For En (fig.41) used 30 µl of 1 U GST solution
Pipetted the samples after the scheme in fig.41

Measurement

Start the reactions with 90 µl master mix and start measurement

Result

All data and graphs of GST - assay 19 and 20 were collected in an Excel file: freiburg_2016_19.09.16_gst-assay_master-1.xlsx

23.09.2016

GST – assay 21

Scheme:

Fig.42: Pipetting scheme for GST – assay with 0.1 U, empty wells and blank measured with PBS instead of GST as control, grey wells were not used.

Prepared

Sample of 0.1 U and master mix: 70 µl CDNB + 70 µl GSH + 560 µl PBS, increased amount of substrates.

Measurement

Start the reaction with 90 µl master mix.
Start the measurement wit settings as before, which are described also in the reader protocol: 23.09.16_gst-assay_01_settings.xlsx

GST – assay 22

Reduced amount of substrates from 70 µl to 50 µl and increased volume from 90 µl to 110 µl used blank of GST – assay 21 (scheme fig.42)
Scheme:

Fig.43: Pipetting scheme for GST – assay 22. 0.1 Units GST for sample wells and as reference empty which contains PBS and master mix.

Prepared

Prepared 0.1 U samples of GST and master mix: 50 µl CDNB + 50 µl GSH + 900 µl PBS
pipetted 30 µl GST and PBS after the scheme (fig.43)

Measurement

Start the reactions with 110 µl master mix
started measurement with setting which are described in the plate reader protocol:23.09.16_gst-assay_02_settings.xlsx

Result: All data and graphs of GST - assay 22 and 23 were collected in an Excel file: freiburg_23.09.16_gst-assay_master-1.xlsx

lab book, labor

26.09.2016

GST assay

with E.coli cell lysate
neg. control: DH5α
pos. control: 0,05 U GST
constructs 131, 134 & 138 (concentrations: 0,05 0,075 0,1 [µg/µl] protein per well)

Didn't work, the concentrations were too low.

29.09.2016

GST assay

concentrations: 0,1 U 0,08 U 0,06 U 0,04 U
0,08 U: 48 µl 5 U/ml stock + 60 µl PBS
0,06 U: 36 µl stock + 72 µl PBS
0,04 U: 24 µl stock + 96 µl PBS
1,7 ml master mix: 85 µl CDNB + 85 µl GSH + 1530 µl PBS
40 µl enzyme solution + 100 µl master mix per well

Didn't work, maybe pipetting error.

30.09.2016

GST assay

E.coli lysate

construct 134
15 µl per well: 30 µl lysate + 150 µl PBS → 69 µg proteins per well
30 µl per well: 60 µl lysate + 120 µl PBS → 138 µg proteins per well
60 µl per well: 120 µl lysate + 60 µl PBS → 276 µg proteins per well
90 µl per well: 180 µl lysate → 414 µg proteins per well
60 µl + GST per well: 120 µl lysate + 0,05 U (20 µl 5 U/ml stock + 40 µl PBS)

DH5α
30 µl per well: 60 µl lysate + 120 µl PBS → 57 µg proteins per well

0,1 U
40 µl 5 U/ml stock + 140 µl PBS

1,5ml master mix: 75 µl CDNB + 75 µl GSH + 1350 µl PBS

90 µl sample + 90 µl master mix per well

30.09.16_gst-assay_01_settings.xlsx

GST assay

E.coli lysate
constructs 131 & 138

131
28 µl → 148,4 µg proteins
52 µl → 275,6 µg proteins
75 µl → 397,5 µg proteins

138
25 µl → 147,8 µg proteins
47 µl → 277,3 µg proteins
68 µl → 401,2 µg proteins

DH5α
79 µl → 150,1 µg proteins

DH5α+
79 µl + 0,05 U (10 µl stock + 1 µl PBS)

0,05 U

1,9 ml master mix: 95 µl CDNB + 95 µl GSH + 1710 µl PBS

90 µl sample + 90 µl master mix

Results

30.09.16_gst-assay_master.xlsx

02.10.2016

GST assay

GST-spores
constructs: Z+GST & WT+GST
concentrations: 50 million, 25 million, 10 million, 5 million spores
stock 100 million spores in 500 µl
50 million: 250 µl stock per well
25 million: 125 µl stock per well + 125 µl PBS
10 million: 50 µl stock per well + 200 µl PBS
5 million: 25 µl stock per well + 225 µl PBS

WT as control: 250 µl per well

0,05 U GST

1,4 ml master mix: 70 µl CDNB + 70 µl GSH + 1260 µl PBS

250 µl sample + 90 µl master mix per well

done again with the CotZ + GST construct and the CotZ construct purified

Results

The spores don't show a difference between GST-construct spores and WT spores or show no activity at all.

03.10.16

GST assay

measured E.coli lysates again with about 150 µg, 275 µg and 400 µg of proteins

Results

The GST spores showed no difference in activity compared to the wild type.

GST assay

measured the WT + GST spores purified
concentrations: 50 million, 25 million, 10 million, 5 million spores
controls: pos. 0,05 U GST & neg. untransformed WT spores

Results

The spores showed no activity.

04.10.16

GST-assay

measured master mix with freshly prepared GSH in ethanol and freshly prepared GSH in demineralized water
400 µl master mix: 20 µl CDNB + 20 µl GSH + 360 µl PBS

Results

The GSH dissolved in EtOH is a little bit more stable.

05.10.16

GST-assay

measured 0,1 U; 0,08 U; 0,06 U; 0,04 U
3 ml master mix: 150 µl CDNB + 150 µl GSH + 2700 µl PBS

Results

There's a measurable difference between the Units, but it's still a slight curve.

05.10.16_gst-assay_0_1u_-_0_04u.xlsx

06.10.16

GST-assay

measured E.coli lysates
DH5α: 6,8 µg/µl proteins
138: 6,8 µg/µl
134: 6,3 µg/µl

concentrations: ~400 µg; ~500 µg; ~600 µg per well

controls: 0,05 U, empty (PBS+master mix)

06.10.16_gst-assay_lysates_settings.xlsx

results see below 10.10.16

GST-assay

measured CotZ-GST knock-out spores and WT spores
50 million - 5 million spores
washed unpurified spores 3 times with PBS

controls:
pos. 0,05 U GST (4*10 µl 5 U/ml stock + 960 µl PBS)
neg. empty (250 µl PBS + 90 µl master mix)
untrans. WT (50-5 million)

3 ml master mix: 150 µl CDNB + 150 µl GSH + 2700 µl PBS

250 µl sample + 90 µl master mix

Results

The spores showed no activity.

08.10.16

Shampoo

tested: aGFPnano-CotZ(159Z) spores

Plate preparation

coated 68 wells with GFP, coating concentration GFP 20 µg/ml and coating volume 100 µl
incubated 2 h at room temerature
washed twice with 200 µl PBS
blocked with 200 µl 5% nonfat dry milk in PBS
incubated for 2 h at roomtemperatur
washed twice with PBS
measured the fluorescence of coated GFP

Washing

used 200 µl washing of solutions:

SDS: 1%, 0.75%, 0.5%, 0.25% (with spores, nanobody and only SDS)
Tween20: 22%, 24%, 26%, 28% (with spores, nanobody and only Tween20)
SH1: 12%, 14%, 16%, 18% (with spores, nanobody and only SH1)
Controls: nanobody in PBS, WT - spore in PBS, construct spore (159Z) in PBS, 2% SDS & PBS

incubated wash solutions on the plate for 5 min
shook for 5 min at 300 rpm
washed twice with PBS
measured the emission of remaining GFP

Results

bba_k2114002_agfpnano_ha_g4s_cotz_in_δcotz_.xlsx

09.10.16

GST-assay

with GST-CotZ(153)-, CotG-GST(156)-, GST-CotG(167)- & CgeA-GST(168)-construct spores.
50 million - 5 million spores

controls:
pos. 0,05 U GST
empty
untrans. WT

3,4 ml master mix: 170 µl CDNB + 170 µl GSH + 3060 µl PBS

09.10.16_gst-assay_settings.xlsx

Results

see below 11.10.16

Shampoo

tested: aGFPnano-CotG(151G) spores

Plate preparation

coated 68 wells with GFP, coating concentration GFP 20 µg/ml and coating volume 100 µl
incubated 2 h at room temerature
washed twice with 200 µl PBS
blocked with 200 µl 5% nonfat dry milk in PBS
incubated for 2 h at roomtemperatur
washed twice with PBS
measured the fluorescence of coated GFP

Washing

used 200 µl washing of solutions:

SDS: 1%, 0.75%, 0.5%, 0.25% (with spores, nanobody and only SDS)
Tween20: 22%, 24%, 26%, 28% (with spores, nanobody and only Tween20)
SH1: 12%, 14%, 16%, 18% (with spores, nanobody and only SH1)
Controls: nanobody in PBS, WT - spore in PBS, construct spore (151G) in PBS, 2% SDS & PBS

incubated wash solutions on the plate for 5 min
shook for 5 min at 300 rpm
washed twice with PBS
measured the emission of remaining GFP

Results

bba_k2114009_agfpnano_ha_ahelix_cotg_in_delta_cotg.xlsx

10.10.16

GST-assay

measured E. coli lysates again with only one concentration (600 µg)
controls:
pos. 0,05 U GST
empty
untrans. WT

Results

06.10.16_10.10.16_gst-assay_lysates_master.xlsx

Shampoo

tested: aGFPnano-CotZ(157Z) spores

Plate preparation

coated 68 wells with GFP, coating concentration GFP 20 µg/ml and coating volume 100 µl
incubated 2 h at room temerature
washed twice with 200 µl PBS
blocked with 200 µl 5% nonfat dry milk in PBS
incubated for 2 h at roomtemperatur
washed twice with PBS
measured the fluorescence of coated GFP

Washing

used 200 µl washing of solutions:

SDS: 1%, 0.75%, 0.5%, 0.25% (with spores, nanobody and only SDS)
Tween20: 22%, 24%, 26%, 28% (with spores, nanobody and only Tween20)
SH1: 12%, 14%, 16%, 18% (with spores, nanobody and only SH1)
Controls: nanobody in PBS, WT - spore in PBS, construct spore (157Z) in PBS, 2% SDS & PBS

incubated wash solutions on the plate for 5 min
shook for 5 min at 300 rpm
washed twice with PBS
measured the emission of remaining GFP

Results

bba_k2114001_agfpnano_ha_ahelix_cotz_in_δcotz_spore.xlsx

11.10.16 Shampoo

prepared 3 plates
tested: aGFPnano-CotZ(159WT), aGFPnano-CotZ(157WT), aGFPnano-CotG(151WT)

Plate preparation

coated 68 wells with GFP, coating concentration GFP 20 µg/ml and coating volume 100 µl
incubated 2 h at room temerature
washed twice with 200 µl PBS
blocked with 200 µl 5% nonfat dry milk in PBS
incubated for 2 h at roomtemperatur
washed twice with PBS
measured the fluorescence of coated GFP

Washing

used 200 µl washing of solutions:

SDS: 1%, 0.75%, 0.5%, 0.25% (with spores, nanobody and only SDS)
Tween20: 22%, 24%, 26%, 28% (with spores, nanobody and only Tween20)
SH1: 12%, 14%, 16%, 18% (with spores, nanobody and only SH1)
Controls: nanobody in PBS, WT - spore in PBS, construct spore in PBS, 2% SDS & PBS

incubated wash solutions on the plate for 5 min
shook for 5 min at 300 rpm
washed twice with PBS
measured the emission of remaining GFP

@@ Line 7: / Line 7: @@
 <head>
+    <!--<link rel="stylesheet" type="text/css" href="style.css">-->
      <style>
          .level3 {
@@ Line 65: / Line 66: @@
          <div class="para_center_20">
              <div class="level1">
-                 <h5 class="sectionedit1">Labbook enzymatic activity and sporeshampoo</h5>
+                 <h5 class="sectionedit1">Group 5 - Enzymatic activity and Sporeshampoo</h5>
-                 <p></p>
+                 <div> The focus of this group lays on the confirmation of the enzymatic activity of our spores. To show that the GST is correctly expressed by the spores a GST-assay is adjusted to the use in a plate reader. Another application of the spores would be for a (anti-dandruff) shampoo, so this is tested by washing a GFP-coated surface with different detergents and our construct and measuring the fluorescence. </div>
              </div>
          </div>
@@ Line 77: / Line 78: @@
              <!-- EDIT2 SECTION "05.08.2016" [59-82] -->
              <h4 class="sectionedit3"><a name="plate_coated_with_gfp" id="plate_coated_with_gfp">plate coated with GFP:</a></h4>
-             <div class="level3">
+             <div class="level3_1">
                  <ol>
                      <li class="level1">
-                         <div class="li"> Prepared coating solution: used Protocol from “Abeam: direct Elisa coating”. Took 0.78g Na2CO3, 1.5g NaHCO3 and dissolve it in 250ml H2O, pH: 9.5.
+                         <div class="li"> Prepared coating solution: used protocol from “Abeam: direct Elisa coating”. Took 0.78 g Na2CO3, 1.5 g NaHCO3 and dissolved it in 250 ml H2O, pH: 9.5.
                              <br/> </div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Dilute stock solution of GFP (1350µg/ml) in coating buffer for coating concentration of 20µg/ml.
+                         <div class="li"> Dilute stock solution of GFP (1350µg/ml) in coating buffer for coating concentration of 20 µg/ml.
                              <br/> </div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Coted 93 wells with 50µl coating solution
+                         <div class="li"> Coated 93 wells with 50µl coating solution
                              <br/> </div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Incubated for 2h at room temperature
+                         <div class="li"> Incubated for 2 h at room temperature
                              <br/> </div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Washed the wells twice with PBS (V=200µl)
+                         <div class="li"> Washed the wells twice with PBS (V=200 µl)
                              <br/> </div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Blocked it with 5% nonfat dry milk in PBS (2.5g in 50ml PBS), blocked coated wells with 200µl solution.
+                         <div class="li"> Blocked it with 5% nonfat dry milk in PBS (2.5 g in 50 ml PBS), blocked coated wells with 200 µl solution.
                              <br/> </div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Incubated for 2h at room temperature.
+                         <div class="li"> Incubated for 2 h at room temperature.
                              <br/> </div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Washed blocked wells twice with PBS (V=200µl)
+                         <div class="li"> Washed blocked wells twice with PBS (V=200 µl)
                              <br/> </div>
                      </li>
@@ Line 115: / Line 116: @@
              <!-- EDIT3 SECTION "plate coated with GFP:" [83-726] -->
              <h4 class="sectionedit4"><a name="measuring" id="measuring">Measuring:</a></h4>
-             <div class="level3">
+             <div class="level3_1">
-                 <p> Measured the emission from top Measured the emission from bottom
+                 <p> Measured the emission from top, Measured the emission from bottom
                      <br/> </p>
              </div>
@@ Line 122: / Line 123: @@
              <h3 class="sectionedit5"><a name="section06082016" id="section06082016">06.08.2016</a></h3>
              <div class="level2">
-                 <p> Measured the plate from 05.08.2016 again with only one Blank:
+                 <p> Measured the plate from 05.08.2016 again with only one blank:
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:freiburg_2016_06.08.2016_flourecence_emisseion_top.png" class="media" title="labor:freiburg_2016_06.08.2016_flourecence_emisseion_top.png"><img src="https://static.igem.org/mediawiki/2016/3/36/T--Freiburg--LabJournal_g5_01.png" class="media" alt="" /></a>
-                     <br/> fig.1: The Average over replicates of fluorescence in RFU. Measured was the fluorescence of coated GFP (20 µg/ml) and the auto fluorescence of empty Wells as a negative control, by measuring from top with the Plate reader.
+                     <br/> fig.1: The Average over replicates of fluorescence in RFU. Measured the fluorescence of coated GFP (20 µg/ml) and the auto fluorescence of empty wells as a negative control, by measuring from top with the plate reader.
                      <br/>
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:freiburg_2016_06.08.2016_flourecence_emisseion_bottom.png" class="media" title="labor:freiburg_2016_06.08.2016_flourecence_emisseion_bottom.png"><img src="https://static.igem.org/mediawiki/2016/f/f6/T--Freiburg--LabJournal_g5_02.png" class="media" alt="" /></a>
-                     <br/> fig.2: The Average over replicates of fluorescence in RFU. Measured was the fluorescence of coated GFP (20 µg/ml) and the auto fluorescence of empty Wells as a negative control, by measuring from bottom with the Plate reader.
+                     <br/> fig.2: The Average over replicates of fluorescence in RFU. Measured the fluorescence of coated GFP (20 µg/ml) and the auto fluorescence of empty wells as a negative control, by measuring from bottom with the Plate reader.
                      <br/>
                      <br/> Prepared coating solutions with several concentrations:
@@ Line 155: / Line 156: @@
              <!-- EDIT6 SECTION "07.08.2016" [1642-1664] -->
              <h4 class="sectionedit7"><a name="coated_med_binding_greiner_plate" id="coated_med_binding_greiner_plate">Coated MED. Binding Greiner Plate:</a></h4>
-             <div class="level3">
+             <div class="level3_1">
-                 <p> <strong>Coat the wells with 50µl from prepared coating solution:</strong>
+                 <p> <strong>Coated the wells with 50 µl from prepared coating solution:</strong>
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:freiburg_2016_06.08.2016_flourecence_emisseion_of_different_coated_gfp_concentrations.png" class="media" title="labor:freiburg_2016_06.08.2016_flourecence_emisseion_of_different_coated_gfp_concentrations.png"><img src="https://static.igem.org/mediawiki/2016/a/ad/T--Freiburg--LabJournal_g5_03.png" class="media" alt="" /></a>
-                     <br/> fig.3: The Average over replicates of fluorescence in RFU. Measured was the fluorescence of coated GFP (20 µg/ml, 28.7 µg/ml, 41.8 µg/ml and 35.6 µg/ml) and the auto fluorescence of empty Wells as a negative control, by measuring from top with the Plate reader.
+                     <br/> fig.3: The Average over replicates of fluorescence in RFU. Measured the fluorescence of coated GFP (20 µg/ml, 28.7 µg/ml, 41.8 µg/ml and 35.6 µg/ml) and the auto fluorescence of empty wells as a negative control, by measuring from top with the Plate reader.
                      <br/>
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:freiburg_2016_06.08.2016_flourecence_emisseion_of_different_coated_gfp_concentrations_anad_gfpin_solution_.png" class="media" title="labor:freiburg_2016_06.08.2016_flourecence_emisseion_of_different_coated_gfp_concentrations_anad_gfpin_solution_.png"><img src="https://static.igem.org/mediawiki/2016/thumb/f/f8/T--Freiburg--LabJournal_g5_04.png/800px-T--Freiburg--LabJournal_g5_04.png" class="media" alt="" /></a>
-                     <br/> fig.4: The Average over replicates of fluorescence in RFU. Measured was the fluorescence of coated GFP (20 µg/ml, 28.7 µg/ml, 41.8 µg/ml and 35.6 µg/ml), the auto fluorescence of empty Wells as a negative control and the fluorescence of GFP (20 µg/ml, 28.7 µg/ml, 41.8 µg/ml and 35.6 µg/ml) in solution as positive control, by measuring from top with the Plate reader.
+                     <br/> fig.4: The Average over replicates of fluorescence in RFU. Measured the fluorescence of coated GFP (20 µg/ml, 28.7 µg/ml, 41.8 µg/ml and 35.6 µg/ml), the auto fluorescence of empty wells as a negative control and the fluorescence of GFP (20 µg/ml, 28.7 µg/ml, 41.8 µg/ml and 35.6 µg/ml) in solution as positive control, by measuring from top with the plate reader.
                      <br/>
                      <br/> </p>
@@ Line 183: / Line 184: @@
                      </li>
                      <li class="level1">
-                         <div class="li"> Spores: WT, B53, B54; Used 50µl/well, for Spores was purified by washing with PBS, to remove the medium.</div>
+                         <div class="li"> Spores: WT, B53, B54; used 50 µl/well, spores were purified by washing with PBS, to remove the medium.</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> used GFP coating concentrations of: 40µg/ml, 45µg/ml, 60µg/ml, 70µg/ml, 80µg/ml, 90µg/ml; used 50µl/well</div>
+                         <div class="li"> used GFP coating concentrations of: 40 µg/ml, 45 µg/ml, 60 µg/ml, 70 µg/ml, 80 µg/ml, 90 µg/ml; used 50 µl/well</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> m.Cherry: 20µg/ml, 40µg/ml, 80µg/ml</div>
+                         <div class="li"> m.Cherry: 20 µg/ml, 40 µg/ml, 80 µg/ml</div>
                      </li>
                  </ol>
                  <p>
-                     <br/> coted over night by 4°C in the fridge.
+                     <br/> coated over night at 4°C in the fridge.
                      <br/>
                      <br/> </p>
@@ Line 202: / Line 203: @@
              <!-- EDIT9 SECTION "09.08.2016" [3056-3078] -->
              <h4 class="sectionedit10"><a name="blocked" id="blocked">Blocked</a></h4>
-             <div class="level3">
+             <div class="level3_1">
                  <ol>
                      <li class="level1">
-                         <div class="li"> Washed plate for previous day twice with 200µl PBS</div>
+                         <div class="li"> Washed plate from previous day twice with 200 µl PBS</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Incubated for 2h at room temperature </div>
+                         <div class="li"> Incubated for 2 h at room temperature </div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Blocked with 200µl 5% nonfat dry milk in PBS</div>
+                         <div class="li"> Blocked with 200 µl 5% nonfat dry milk in PBS</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Incubated for 2h at room temperature</div>
+                         <div class="li"> Incubated for 2 h at room temperature</div>
                      </li>
                  </ol>
@@ Line 222: / Line 223: @@
              <!-- EDIT10 SECTION "Blocked" [3079-3289] -->
              <h4 class="sectionedit11"><a name="measured" id="measured">Measured</a></h4>
-             <div class="level3">
+             <div class="level3_1">
-                 <p> Measured the Fluorescents at two different focal heights:
+                 <p> Measured the fluorescence at two different focal heights:
                      <br/> <strong>22mm:</strong>
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:freiburg_2016_09.08.2016average_of_the_rfu_focal_height_22mm_spore_and_gef_coated.png" class="media" title="labor:freiburg_2016_09.08.2016average_of_the_rfu_focal_height_22mm_spore_and_gef_coated.png"><img src="https://static.igem.org/mediawiki/2016/thumb/8/8b/T--Freiburg--LabJournal_g5_05.png/800px-T--Freiburg--LabJournal_g5_05.png" class="media" alt="" /></a>
-                     <br/> fig.5: Average of fluorescence over replicates of samples (B53_coated 18mill., B53_solution 18mill., B53_coated 9mill., B53_solution 9mill., B53_coated 1.8mill., B53_solution 1.8mill., B54_cated 18mill., B54_solution 18mill., B54_coated 9mill., B54_solution 9mill., B54_coated 1.8mill., B54_solution 1.8mill., GFP_coated 20µg/ml, GFP_coated 35.6µg/ml, GFP_coated 40µg/ml, GFP_coated 45µg/ml, GFP_coated 60Mg/ml, GFP_coated 70µg/ml, GFP_coated 80µg/ml, GFP_coated 90µg/ml, m.Cherry_coated 20µg/ml, m.Cherry_coated 40µg/ml, m.Cherry_coated 80µg/ml, WT_coated 18mill.) and an empty well as negative control, at a focal height of 22mm.
+                     <br/> fig.5: Average of fluorescence over replicates of samples (B53_coated 18 mill., B53_solution 18 mill., B53_coated 9 mill., B53_solution 9 mill., B53_coated 1.8 mill., B53_solution 1.8 mill., B54_cated 18 mill., B54_solution 18 mill., B54_coated 9 mill., B54_solution 9 mill., B54_coated 1.8 mill., B54_solution 1.8 mill., GFP_coated 20 µg/ml, GFP_coated 35.6 µg/ml, GFP_coated 40 µg/ml, GFP_coated 45 µg/ml, GFP_coated 60 µg/ml, GFP_coated 70 µg/ml, GFP_coated 80 µg/ml, GFP_coated 90 µg/ml, m.Cherry_coated 20 µg/ml, m.Cherry_coated 40 µg/ml, m.Cherry_coated 80 µg/ml, WT_coated 18 mill.) and an empty well as negative control, at a focal height of 22 mm.
                      <br/>
-                     <br/> <strong>2mm:</strong>
+                     <br/> <strong>2 mm:</strong>
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:freiburg_2016_09.08.2016average_of_the_rfu_focal_height_2mm_spore_and_gef_coated.png" class="media" title="labor:freiburg_2016_09.08.2016average_of_the_rfu_focal_height_2mm_spore_and_gef_coated.png"><img src="https://static.igem.org/mediawiki/2016/thumb/d/db/T--Freiburg--LabJournal_g5_06.png/800px-T--Freiburg--LabJournal_g5_06.png" class="media" alt="" /></a>
-                     <br/> fig.6: Average of fluorescence over replicates of samples (B53_coated 18mill., B53_solution 18mill., B53_coated 9mill., B53_solution 9mill., B53_coated 1.8mill., B53_solution 1.8mill., B54_cated 18mill., B54_solution 18mill., B54_coated 9mill., B54_solution 9mill., B54_coated 1.8mill., B54_solution 1.8mill., GFP_coated 20µg/ml, GFP_coated 35.6µg/ml, GFP_coated 40µg/ml, GFP_coated 45µg/ml, GFP_coated 60Mg/ml, GFP_coated 70µg/ml, GFP_coated 80µg/ml, GFP_coated 90µg/ml, m.Cherry_coated 20µg/ml, m.Cherry_coated 40µg/ml, m.Cherry_coated 80µg/ml, WT_coated 18mill.) and an empty well as negative control, at a focal height of 2mm.
+                     <br/> fig.6: Average of fluorescence over replicates of samples (B53_coated 18 mill., B53_solution 18 mill., B53_coated 9 mill., B53_solution 9 mill., B53_coated 1.8 mill., B53_solution 1.8 mill., B54_cated 18 mill., B54_solution 18 mill., B54_coated 9 mill., B54_solution 9 mill., B54_coated 1.8 mill., B54_solution 1.8 mill., GFP_coated 20 µg/ml, GFP_coated 35.6 µg/ml, GFP_coated 40 µg/ml, GFP_coated 45 µg/ml, GFP_coated 60 µg/ml, GFP_coated 70 µg/ml, GFP_coated 80 µg/ml, GFP_coated 90 µg/ml, m.Cherry_coated 20 µg/ml, m.Cherry_coated 40 µg/ml, m.Cherry_coated 80 µg/ml, WT_coated 18 mill.) and an empty well as negative control, at a focal height of 2 mm.
                      <br/>
                      <br/> </p>
@@ Line 247: / Line 248: @@
              <!-- EDIT12 SECTION "13.08.2016" [4898-4920] -->
              <h4 class="sectionedit13"><a name="dried_in_pbs" id="dried_in_pbs">Dried in PBS</a></h4>
-             <div class="level3">
+             <div class="level3_1">
                  <p> Scheme:
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:freiburg_2016_dry_experiment_scheme.png" class="media" title="labor:freiburg_2016_dry_experiment_scheme.png"><img src="https://static.igem.org/mediawiki/2016/7/76/T--Freiburg--LabJournal_g5_07.png" class="media" alt="" /></a>
-                     <br/> fig.7: Pipette scheme for the measuring of fluorescence at the surface by several concentrations of GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml) and two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54). The shaded wells shown dried wells with PBS, were the solution of GFP/Spores was dried by 37°C in the incubator.
+                     <br/> fig.7: Pipetting scheme for the measuring of fluorescence at the surface with several concentrations of GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml) and two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54). The shaded wells show wells with PBS dried, where the solution of GFP/spores was dried at 37°C in the incubator.
                      <br/>
                      <br/> </p>
                  <ol>
                      <li class="level1">
-                         <div class="li"> took from B54 and B53 aliquot 108µl (108 mill. Spores) and 54µl (54 mill. Spores)
+                         <div class="li"> took from B54 and B53 aliquot 108 µl (108 mill. Spores) and 54µl (54 mill. spores)
                              <br/> </div>
                      </li>
                      <li class="level1">
-                         <div class="li"> washed them by spinning down at 13000rct for 13min and resuspend the pellet in PBS, three times</div>
+                         <div class="li"> washed them by spinning down at 13000rpm for 13min and resuspended the pellet in PBS, repeated three times</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> resuspend final pellet in 300µl PBS -pipetted 50µl solution into the shaded wells (scheme) </div>
+                         <div class="li"> resuspended final pellet in 300µl PBS -pipetted 50 µl solution into the shaded wells (scheme) </div>
                      </li>
                      <li class="level1">
-                         <div class="li"> incubated at 37°C for 5,5h
+                         <div class="li"> incubated at 37°C for 5,5 h
                              <br/> </div>
                      </li>
@@ Line 275: / Line 276: @@
              <h5><a name="measured1" id="measured1">Measured</a></h5>
              <div class="level4">
-                 <p> Measures the plate by different focal height and gain values:
+                 <p> Measured the plate with different focal heights and gain values:
                      <br/> </p>
                  <ul>
                      <li class="level1">
-                         <div class="li"> Focal height 22mm, gain 1740</div>
+                         <div class="li"> Focal height 22 mm, gain 1740</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Focal height self-adjusted at 3.6mm, gain 1740</div>
+                         <div class="li"> Focal height self-adjusted at 3.6 mm, gain 1740</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Focal height 3.6mm, gain 2960</div>
+                         <div class="li"> Focal height 3.6 mm, gain 2960</div>
                      </li>
                  </ul>
@@ Line 297: / Line 298: @@
                  <p>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:freiburg_2016_13.08.2016dry_01_focal22_gain25_barchart.png" class="media" title="labor:freiburg_2016_13.08.2016dry_01_focal22_gain25_barchart.png"><img src="https://static.igem.org/mediawiki/2016/thumb/f/f2/T--Freiburg--LabJournal_g5_08.png/800px-T--Freiburg--LabJournal_g5_08.png" class="media" alt="" /></a>
-                     <br/> fig.8: Average of relative fluorescence over Replicates of dried in PBS, GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml), two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54) and empty wells as negative control. It was used a setting with a focal height of 22mm and gain value of 1740.
+                     <br/> fig.8: Average of relative fluorescence over replicates of dried in PBS, GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml), two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54) and empty wells as negative control. Used setting: focal height of 22 mm and gain value of 1740.
                      <br/>
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:freiburg_2016_13.08.2016_dry_01_focaladjustment-self3.6_gain25_barchart.png" class="media" title="labor:freiburg_2016_13.08.2016_dry_01_focaladjustment-self3.6_gain25_barchart.png"><img src="https://static.igem.org/mediawiki/2016/thumb/f/f2/T--Freiburg--LabJournal_g5_08.png/800px-T--Freiburg--LabJournal_g5_08.png" class="media" alt="" /></a>
-                     <br/> fig.9: Average of relative fluorescence over Replicates of dried in PBS, GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml), two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54) and empty wells as negative control. It was used a setting with a focal height of 3.6mm and gain value of 1740.
+                     <br/> fig.9: Average of relative fluorescence over replicates of dried in PBS, GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml), two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54) and empty wells as negative control. Used setting: focal height of 3.6 mm and gain value of 1740.
                      <br/>
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:freiburg_2016_13.08.2016_dry_01_focal3.6_gain2960_barchart.png" class="media" title="labor:freiburg_2016_13.08.2016_dry_01_focal3.6_gain2960_barchart.png"><img src="https://static.igem.org/mediawiki/2016/thumb/f/f3/T--Freiburg--LabJournal_g5_10.png/800px-T--Freiburg--LabJournal_g5_10.png" class="media" alt="" /></a>
-                     <br/> fig.10: Average of relative fluorescence over Replicates of dried in PBS, GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml), two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54) and empty wells as negative control. It was used a setting with a focal height of 3.6mm and gain value of 2960.
+                     <br/> fig.10: Average of relative fluorescence over replicates of dried in PBS, GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml), two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54) and empty wells as negative control. Used setting: focal height of 3.6 mm and gain value of 2960.
                      <br/>
                      <br/> </p>
@@ Line 313: / Line 314: @@
                  <p>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:freiburg_2016_13.08.2016_fluorescein_wet_01_focal22_gain1740_barchart.png" class="media" title="labor:freiburg_2016_13.08.2016_fluorescein_wet_01_focal22_gain1740_barchart.png"><img src="https://static.igem.org/mediawiki/2016/thumb/d/d8/T--Freiburg--LabJournal_g5_11.png/800px-T--Freiburg--LabJournal_g5_11.png" class="media" alt="" /></a>
-                     <br/> fig.11: Average of relative fluorescence over Replicates with PBS on top GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml), two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54) and empty wells as negative control. It was used a setting with a focal height of 22mm and gain value of 1740.
+                     <br/> fig.11: Average of relative fluorescence over replicates with PBS on top GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml), two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54) and empty wells as negative control. It was used a setting with a focal height of 22 mm and gain value of 1740.
                      <br/>
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:freiburg_2016_13.08.2016_fluorescein_wet_01_focal3.6_gain1740_barchart.png" class="media" title="labor:freiburg_2016_13.08.2016_fluorescein_wet_01_focal3.6_gain1740_barchart.png"><img src="https://static.igem.org/mediawiki/2016/thumb/3/3c/T--Freiburg--LabJournal_g5_12.png/800px-T--Freiburg--LabJournal_g5_12.png" class="media" alt="" /></a>
-                     <br/> fig.12: Average of relative fluorescence over Replicates with PBS on top GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml), two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54) and empty wells as negative control. It was used a setting with a focal height of 3.6mm and gain value of 1740.
+                     <br/> fig.12: Average of relative fluorescence over replicates with PBS on top GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml), two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54) and empty wells as negative control. It was used a setting with a focal height of 3.6 mm and gain value of 1740.
                      <br/>
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:freiburg_2016_13.08.2016_fluorescein_wet_01_focal3.6_gain2960_barchart.png" class="media" title="labor:freiburg_2016_13.08.2016_fluorescein_wet_01_focal3.6_gain2960_barchart.png"><img src="https://static.igem.org/mediawiki/2016/thumb/4/46/T--Freiburg--LabJournal_g5_13.png/800px-T--Freiburg--LabJournal_g5_13.png" class="media" alt="" /></a>
-                     <br/> fig.13: Average of relative fluorescence over Replicates with PBS on top GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml), two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54) and empty wells as negative control. It was used a setting with a focal height of 3.6mm and gain value of 2960.
+                     <br/> fig.13: Average of relative fluorescence over replicates with PBS on top GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml), two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54) and empty wells as negative control. Used setting: focal height of 3.6 mm and gain value of 2960.
                      <br/>
                      <br/> </p>
@@ Line 336: / Line 337: @@
              <!-- EDIT14 SECTION "14.08.2016 – 15.08.2016" [8433-8470] -->
              <h4 class="sectionedit15"><a name="dried_in_h2o" id="dried_in_h2o">Dried in H2O</a></h4>
-             <div class="level3">
+             <div class="level3_1">
                  <p> scheme:
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:freiburg_2016_dry_experiment_scheme.png" class="media" title="labor:freiburg_2016_dry_experiment_scheme.png"><img src="https://static.igem.org/mediawiki/2016/thumb/7/76/T--Freiburg--LabJournal_g5_07.png/120px-T--Freiburg--LabJournal_g5_07.png" class="media" alt="" /></a>
-                     <br/> fig.14: Pipette scheme for the measuring of fluorescence at the surface by several concentrations of GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml) and two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54). The shaded wells shown dried wells with H2O, were the solution of GFP/Spores was dried by 37°C in the incubator.
+                     <br/> fig.14: Pipette scheme for the measuring of fluorescence at the surface by several concentrations of GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml) and two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54). The shaded wells shown dried wells with H2O, were the solution of GFP/spores was dried by 37°C in the incubator.
                      <br/>
                      <br/> </p>
@@ Line 348: / Line 349: @@
                  <ul>
                      <li class="level1">
-                         <div class="li"> Prepared dilution series of GFP and Spores in H2O as in the scheme</div>
+                         <div class="li"> Prepared dilution series of GFP and spores in H2O as in the scheme</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Pipetted 100µl per well and incubated it at 37°C for 12h 40min.</div>
+                         <div class="li"> Pipetted 100 µl per well and incubated it at 37°C for 12 h 40 min.</div>
                      </li>
                  </ul>
@@ Line 360: / Line 361: @@
              <div class="level4">
                  <p> Measured the plate with three different settings:
-                     <br/> * Focal height 22mm, gain 1740 * Focal height 3.6mm, gain 1740 * Focal height 3.6mm, gain 2960
+                     <br/> * Focal height 22 mm, gain 1740 * Focal height 3.6 mm, gain 1740 * Focal height 3.6 mm, gain 2960
                      <br/> </p>
              </div>
              <h5><a name="result1" id="result1">Result:</a></h5>
              <div class="level4">
-                 <p> Focal height 22mm, gain 1740:
+                 <p> Focal height 22 mm, gain 1740:
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:fluorescencewet02_22_1740.png" class="media" title="labor:fluorescencewet02_22_1740.png"><img src="https://static.igem.org/mediawiki/2016/c/cc/T--Freiburg--LabJournal_g5_15.png" class="media" alt="" /></a>
-                     <br/> fig.15: Average of relative fluorescence over Replicates of dried in H2O, GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml), two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54) and empty wells as negative control. It was used a setting with a focal height of 22mm and gain value of 1740.
+                     <br/> fig.15: Average of relative fluorescence over replicates of dried in H2O, GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml), two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54) and empty wells as negative control. Used setting: focal height of 22 mm and gain value of 1740.
                      <br/>
-                     <br/> Focal height 3.6mm, gain 1740:
+                     <br/> Focal height 3.6 mm, gain 1740:
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:fluorescencewet02_focal3.6_gain1740.png" class="media" title="labor:fluorescencewet02_focal3.6_gain1740.png"><img src="https://static.igem.org/mediawiki/2016/thumb/8/83/T--Freiburg--LabJournal_g5_16.png/800px-T--Freiburg--LabJournal_g5_16.png" class="media" alt="" /></a>
-                     <br/> fig.16: Average of relative fluorescence over Replicates of dried in H2O, GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml), two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54) and empty wells as negative control. It was used a setting with a focal height of 3.6mm and gain value of 1740.
+                     <br/> fig.16: Average of relative fluorescence over replicates of dried in H2O, GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml), two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54) and empty wells as negative control. It was used a setting with a focal height of 3.6 mm and gain value of 1740.
                      <br/>
-                     <br/> Focal height 3.6mm, gain 2960:
+                     <br/> Focal height 3.6 mm, gain 2960:
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:freiburg_2016_14.08.2016_fluorescein_wet02_focal3_6_gain2960_barchart.png" class="media" title="labor:freiburg_2016_14.08.2016_fluorescein_wet02_focal3_6_gain2960_barchart.png"><img src="https://static.igem.org/mediawiki/2016/thumb/6/6a/T--Freiburg--LabJournal_g5_17.png/800px-T--Freiburg--LabJournal_g5_17.png" class="media" alt="" /></a>
-                     <br/> fig.17: Average of relative fluorescence over Replicates of dried in H2O, GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml), two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54) and empty wells as negative control. It was used a setting with a focal height of 3.6mm and gain value of 2960.
+                     <br/> fig.17: Average of relative fluorescence over replicates of dried in H2O, GFP (20, 15, 10, 5, 1, 0,1, 0,01, 0,001 µg/ml), two different amounts (18 mill., 9 mill.) of spores of two different types (B53, B54) and empty wells as negative control. Used setting: focal height of 3.6 mm and gain value of 2960.
                      <br/>
                      <br/> </p>
@@ Line 387: / Line 388: @@
              <!-- EDIT16 SECTION "16.08.2016 – 15. 08.2016" [10558-10596] -->
              <h4 class="sectionedit17"><a name="coated" id="coated">Coated</a></h4>
-             <div class="level3">
+             <div class="level3_1">
                  <ul>
                      <li class="level1">
@@ Line 393: / Line 394: @@
                      </li>
                      <li class="level1">
-                         <div class="li"> Coated concentrations of: 20µg/ml (D5 – D7), 15µg/ml (D8 – D10), 10µg/ml (D11, D12 and H5), 5µg/ml (H6 – H8)</div>
+                         <div class="li"> Coated concentrations of: 20 µg/ml (D5 – D7), 15 µg/ml (D8 – D10), 10 µg/ml (D11, D12 and H5), 5 µg/ml (H6 – H8)</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Coated with 50µl </div>
+                         <div class="li"> Coated with 50 µl </div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Incubated for 2h at room temperature</div>
+                         <div class="li"> Incubated for 2 h at room temperature</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Washed wells twice with 200µl PBS</div>
+                         <div class="li"> Washed wells twice with 200 µl PBS</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Blocked with 200µl 5% nonfat dry milk in PBS</div>
+                         <div class="li"> Blocked with 200 µl 5% nonfat dry milk in PBS</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Incubated for 2h at room temperature</div>
+                         <div class="li"> Incubated for 2 h at room temperature</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Washed the wells twice with 200µl PBS</div>
+                         <div class="li"> Washed the wells twice with 200 µl PBS</div>
                      </li>
                  </ul>
@@ Line 419: / Line 420: @@
              <!-- EDIT17 SECTION "Coated" [10597-11090] -->
              <h4 class="sectionedit18"><a name="measured3" id="measured3">Measured</a></h4>
-             <div class="level3">
+             <div class="level3_1">
                  <p> Measured the emission of the fluorescence with several settings:
-                     <br/> Focal height 22mm, gain 1740
+                     <br/> Focal height 22 mm, gain 1740
-                     <br/> Focal height 3.6mm, gain 174
+                     <br/> Focal height 3.6 mm, gain 174
-                     <br/> Focal height 3.6mm, gain 2960
+                     <br/> Focal height 3.6 mm, gain 2960
                      <br/>
                      <br/> </p>
@@ Line 429: / Line 430: @@
              <!-- EDIT18 SECTION "Measured" [11091-11273] -->
              <h4 class="sectionedit19"><a name="result2" id="result2">Result</a></h4>
-             <div class="level3">
+             <div class="level3_1">
-                 <p> Focal height 22mm, gain 1740:
+                 <p> Focal height 22 mm, gain 1740:
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:freiburg_2016_16.08.2016_gfp_coated_focal22_gain_1740.png" class="media" title="labor:freiburg_2016_16.08.2016_gfp_coated_focal22_gain_1740.png"><img src="https://static.igem.org/mediawiki/2016/2/22/T--Freiburg--LabJournal_g5_18.png" class="media" alt="" /></a>
-                     <br/> fig.18: Measured the relative fluorescence over replicates of coated GFP (20µg/ml, 15µg/ml, 10µg/ml, 5µg/ml) and empty wells as negative control, at focal height 22mm with a gain value of 1740.
+                     <br/> fig.18: Measured the relative fluorescence over replicates of coated GFP (20 µg/ml, 15 µg/ml, 10 µg/ml, 5 µg/ml) and empty wells as negative control, at focal height 22 mm with a gain value of 1740.
                      <br/>
-                     <br/> Focal height 3.6mm, gain 1740:
+                     <br/> Focal height 3.6 mm, gain 1740:
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:freiburg_2016_16.08.2016_gfp_coated_focal3.6_gain_1740.png" class="media" title="labor:freiburg_2016_16.08.2016_gfp_coated_focal3.6_gain_1740.png"><img src="https://static.igem.org/mediawiki/2016/0/04/T--Freiburg--LabJournal_g5_19.png" class="media" alt="" /></a>
-                     <br/> fig.19: Measured the relative fluorescence over replicates of coated GFP (20µg/ml, 15µg/ml, 10µg/ml, 5µg/ml) and empty wells as negative control, at focal height 3.6mm with a gain value of 1740.
+                     <br/> fig.19: Measured the relative fluorescence over replicates of coated GFP (20 µg/ml, 15 µg/ml, 10 µg/ml, 5 µg/ml) and empty wells as negative control, at focal height 3.6 mm with a gain value of 1740.
                      <br/>
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:freiburg_2016_16.08.2016_gfp_coated_focal3.6_gain_2960.png" class="media" title="labor:freiburg_2016_16.08.2016_gfp_coated_focal3.6_gain_2960.png"><img src="https://static.igem.org/mediawiki/2016/0/0a/T--Freiburg--LabJournal_g5_20.png" class="media" alt="" /></a>
-                     <br/> fig.20: Measured the relative fluorescence over replicates of coated GFP (20µg/ml, 15µg/ml, 10µg/ml, 5µg/ml) and empty wells as negative control, at focal height 3.6mm with a gain value of 2960.
+                     <br/> fig.20: Measured the relative fluorescence over replicates of coated GFP (20 µg/ml, 15 µg/ml, 10 µg/ml, 5 µg/ml) and empty wells as negative control, at focal height 3.6 mm with a gain value of 2960.
                      <br/>
                      <br/> </p>
@@ Line 451: / Line 452: @@
              <!-- EDIT20 SECTION "19.08.2016" [12206-12228] -->
              <h4 class="sectionedit21"><a name="plate_coated_and_washed" id="plate_coated_and_washed">Plate coated and washed</a></h4>
-             <div class="level3"> </div>
+             <div class="level3_1"> </div>
              <h5><a name="coated1" id="coated1">Coated</a></h5>
              <div class="level4">
@@ Line 459: / Line 460: @@
                      </li>
                      <li class="level1">
-                         <div class="li"> Coating concentration of GFP20µg/ml</div>
+                         <div class="li"> Coating concentration of GFP 20 µg/ml</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Coating volume 50µl</div>
+                         <div class="li"> Coating volume 50 µl</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Incubated for 4h at room temperature</div>
+                         <div class="li"> Incubated for 4 h at room temperature</div>
                      </li>
                  </ol>
@@ Line 475: / Line 476: @@
                  <ol>
                      <li class="level1">
-                         <div class="li"> Washed coated wells twice with 200µ PBS</div>
+                         <div class="li"> Washed coated wells twice with 200 µl PBS</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Blocked with 200µl 5% nonfat dry milk in PBS </div>
+                         <div class="li"> Blocked with 200 µl 5% nonfat dry milk in PBS </div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Incubated for 2h at room temperature</div>
+                         <div class="li"> Incubated for 2 h at room temperature</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Washed wells twice with 200µl PBS</div>
+                         <div class="li"> Washed wells twice with 200 µl PBS</div>
                      </li>
                  </ol>
@@ Line 492: / Line 493: @@
              <h5><a name="measured4" id="measured4">Measured</a></h5>
              <div class="level4">
-                 <p> Measured with a focal height of 3.6mm and a gain value of 1740
+                 <p> Measured with a focal height of 3.6 mm and a gain value of 1740
                      <br/>
                      <br/> </p>
@@ Line 498: / Line 499: @@
              <h5><a name="result3" id="result3">Result</a></h5>
              <div class="level4">
-                 <p> fig.20: Average of relative fluorescence units over replicates of coated GFP (20µg/ml) and empty wells as negative control.
+                 <p> fig.20: Average of relative fluorescence units over replicates of coated GFP (20 µg/ml) and empty wells as negative control.
                      <br/>
                      <br/> </p>
@@ Line 519: / Line 520: @@
                  <ol>
                      <li class="level1">
-                         <div class="li"> Washed the plate after the scheme with 200µl detergent and shook for 2min, pipetted out.</div>
+                         <div class="li"> Washed the plate after the scheme with 200 µl detergent and shook for 2 min, pipetted out.</div>
                      </li>
                      <li class="level1">
@@ Line 531: / Line 532: @@
                      </li>
                      <li class="level1">
-                         <div class="li"> Washed again with 200µl detergent and shook for 2min, pipetted out </div>
+                         <div class="li"> Washed again with 200 µl detergent and shook for 2 min, pipetted out </div>
                      </li>
                      <li class="level1">
@@ Line 580: / Line 581: @@
              <!-- EDIT22 SECTION "21.08.2016" [14376-14398] -->
              <h4 class="sectionedit23"><a name="washing_assay_02" id="washing_assay_02">Washing – Assay 02</a></h4>
-             <div class="level3">
+             <div class="level3_1">
-                 <p> <strong>Prepared different concentrations of detergent:</strong>
+                 <p> <strong>Prepared different concentrations of detergents:</strong>
                      <br/> </p>
                  <ul>
@@ Line 602: / Line 603: @@
                  <p>
                      <br/> For washing used plate and scheme from 19.06.2016 with different concentrations.
-                     <br/> washed the coated wells with the detergents, pipetted 200µl detergent and shook for 2min, pipetted out.
+                     <br/> washed the coated wells with the detergents, pipetted 200 µl detergent and shook for 2 min, pipetted out.
                      <br/> Measured the fluorescence emission after washing with detergent.
                      <br/> Measured the fluorescence emission after washing with PBS.
@@ Line 617: / Line 618: @@
              <h5><a name="results" id="results">Results</a></h5>
              <div class="level4">
-                 <p> fig.24: RFU difference between last measurement of the plate (washed again with PBS) and washed with detergents: SDS (2%; 1,5%; 1%; 0,5%; 0,1%), Triton: 1,5%; 1%; 0,5%; 0,1%), Tween (1,5%; 1%; 0,5%; 0,1%), Shampoo 1 (1,5%; 1%; 0,5%; 0,1%), Shampoo 2 (1,5%; 1%; 0,5%; 0,1%), SDS 2% was used as positive control. Used a focal height of 3.6mm and a gain value of 1740.
+                 <p> fig.24: RFU difference between last measurement of the plate (washed again with PBS) and washed with detergents: SDS (2%; 1,5%; 1%; 0,5%; 0,1%), Triton: 1,5%; 1%; 0,5%; 0,1%), Tween (1,5%; 1%; 0,5%; 0,1%), Shampoo 1 (1,5%; 1%; 0,5%; 0,1%), Shampoo 2 (1,5%; 1%; 0,5%; 0,1%), SDS 2% was used as positive control. Used a focal height of 3.6 mm and a gain value of 1740.
                      <br/>
                      <br/> </p>
@@ Line 632: / Line 633: @@
              <!-- EDIT24 SECTION "23.08.2016" [15447-15469] -->
              <h4 class="sectionedit25"><a name="gst-assay_01" id="gst-assay_01">GST-Assay 01</a></h4>
-             <div class="level3"> </div>
+             <div class="level3_1"> </div>
              <h5><a name="preparation1" id="preparation1">Preparation</a></h5>
              <div class="level4">
-                 <p> <strong>Prepared dilution of GST-Stock (5Units/ml):</strong>
+                 <p> <strong>Prepared dilution of GST stock (5 Units/ml):</strong>
                      <br/> </p>
                  <ul>
                      <li class="level1">
-                         <div class="li"> 1U/ml</div>
+                         <div class="li"> 1 U/ml</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> 0.75U/ml</div>
+                         <div class="li"> 0.75 U/ml</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> 0.5U/ml</div>
+                         <div class="li"> 0.5 U/ml</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> 0.1U/ml</div>
+                         <div class="li"> 0.1 U/ml</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> 0.02U/ml</div>
+                         <div class="li"> 0.02 U/ml</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> 0.001U/ml
+                         <div class="li"> 0.001 U/ml
                              <br/> </div>
                      </li>
                  </ul>
-                 <p> Volume assay cocktail 1.5ml (15µlCDNB [100mM] + 15µl GSH [100mM] + 1470µl PBS).
+                 <p> Volume master mix 1.5 ml: (15 µlCDNB [100 mM] + 15 µl GSH [100 mM] + 1470 µl PBS).
                      <br/>
                      <br/> Plate scheme:
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:freiburg_2016_gst_1_scheme.png" class="media" title="labor:freiburg_2016_gst_1_scheme.png"><img src="https://static.igem.org/mediawiki/2016/9/92/T--Freiburg--LabJournal_g5_25.png" class="media" alt="" /></a>
-                     <br/> fig.25: scheme for measuring the activity of different GST concentrations (1U/ml, 0.75U/ml, 0.5U/ml, 0.1U/ml, 0.02U/ml, 0.001U/ml), Empty (PBS and Assay cocktail), Blank (PBS and Assay cocktail).
+                     <br/> fig.25: scheme for measuring the activity of different GST concentrations (1 U/ml, 0.75 U/ml, 0.5 U/ml, 0.1 U/ml, 0.02 U/ml, 0.001 U/ml), Empty (PBS and master mix), blank (PBS and master mix).
                      <br/>
                      <br/> </p>
@@ Line 669: / Line 670: @@
              <h5><a name="measured6" id="measured6">Measured</a></h5>
              <div class="level4">
-                 <p> start the reaction with 30µl GST as in the scheme (Abb.27), in blank and empty (control) was added 30µl PBS.
+                 <p> start the reaction with 30 µl GST as in the scheme (Abb.27), in blank and empty (control) 30 µl PBS was added.
                      <br/> Setrtings:<a href="https://static.igem.org/mediawiki/2016/f/f7/T--Freiburg--LabJournal_g5_xlsx05.xlsx" class="media mediafile mf_xlsx" title="labor:protocol_gst_run_1_.xlsx">protocol_gst_run_1_.xlsx</a>
                      <br/>
@@ Line 680: / Line 681: @@
                      <br/> </p>
              </div>
-             <!-- EDIT25 SECTION "GST-Assay 01" [15470-16291] -->
+             <!-- EDIT25 SECTION "GST - assay 01" [15470-16291] -->
              <h4 class="sectionedit26"><a name="gst-assay_02" id="gst-assay_02">GST-Assay 02</a></h4>
-             <div class="level3"> </div>
+             <div class="level3_1"> </div>
              <h5><a name="preparation2" id="preparation2">Preparation</a></h5>
              <div class="level4">
@@ Line 689: / Line 690: @@
                  <ul>
                      <li class="level1">
-                         <div class="li"> 0.3U (2x 0.15U) 0.15U (90µl of 5U/ml stock)</div>
+                         <div class="li"> 0.3 U (2x 0.15 U) 0.15 U (90 µl of 5 U/ml stock)</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> 0.25U (2x 0.125U) 0.125U (25µl of 5U/ml stock + 5µl PBS)</div>
+                         <div class="li"> 0.25 U (2x 0.125 U) 0.125 U (25 µl of 5 U/ml stock + 5 µl PBS)</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> 0.2U (2x 0.1U) 0.1U (20µl of 5U/ml stock + 10µl PBS)</div>
+                         <div class="li"> 0.2 U (2x 0.1 U) 0.1 U (20 µl of 5 U/ml stock + 10 µl PBS)</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> 0.15U (2x 0.075U) 0.075U (15µl of 5U/ml stock + 15µl PBS)</div>
+                         <div class="li"> 0.15 U (2x 0.075 U) 0.075 U (15 µl of 5 U/ml stock + 15 µl PBS)</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> 0.1U (2x 0.05U) 0.05U (10µl of 5U/ml stock + 20µl PBS)</div>
+                         <div class="li"> 0.1 U (2x 0.05 U) 0.05 U (10 µl of 5 U/ml stock + 20 µl PBS)</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> 0.05U (2x 0.025U) 0.025U (5µl of 5U/ml stock + 25µl PBS)</div>
+                         <div class="li"> 0.05 U (2x 0.025 U) 0.025 U (5 µl of 5 U/ml stock + 25 µl PBS)</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> 0.02U (2x 0.01U) 0.01U (2µl of 5U/ml stock + 28µl PBS)</div>
+                         <div class="li"> 0.02 U (2x 0.01 U) 0.01 U (2 µl of 5 U/ml stock + 28 µl PBS)</div>
                      </li>
                  </ul>
                  <p>
-                     <br/> volume auf each sample contains 30µl.
+                     <br/> volume of each sample contains 30 µl.
-                     <br/> Prepared assay cocktail (1470µl PBS + 15µl GSH [100mM] + 15µl CDNB [100mM]).
+                     <br/> Prepared mester mix (1470 µl PBS + 15 µl GSH [100 mM] + 15 µl CDNB [100 mM]).
-                     <br/> Pipetted GST – samples after the scheme (fig.26)
+                     <br/> Pipetted GST samples like in the scheme (fig.26)
                      <br/>
                      <br/> Scheme:
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:freiburg_2016_gst_2_scheme.png" class="media" title="labor:freiburg_2016_gst_2_scheme.png"><img src="https://static.igem.org/mediawiki/2016/9/99/T--Freiburg--LabJournal_g5_26.png" class="media" alt="" /></a>
-                     <br/> fig.26: pipetting scheme of GST-amount (0.15U, 0.125U, 0.1U, 0.075U, 0.05U, 0.025U, 0.01U), white empty (Assay cocktail and PBS) in black Blank (Assay cocktail and PBS).
+                     <br/> fig.26: pipetting scheme of GST amount (0.15 U, 0.125 U, 0.1 U, 0.075 U, 0.05 U, 0.025 U, 0.01 U), white=empty (master mix and PBS) and black=blank (master mix and PBS).
                      <br/> </p>
              </div>
@@ Line 725: / Line 726: @@
                  <ul>
                      <li class="level1">
-                         <div class="li"> Started the reaction with 90µl assay cocktail in each well</div>
+                         <div class="li"> Started the reaction with 90 µl master mix in each well</div>
                      </li>
                      <li class="level1">
@@ Line 740: / Line 741: @@
              <h5><a name="results1" id="results1">Results</a></h5>
              <div class="level4">
-                 <p> All data and Graphs were collected in an excel file:<a href="https://static.igem.org/mediawiki/2016/0/05/T--Freiburg--LabJournal_g5_xlsx08.xlsx" class="media mediafile mf_xlsx" title="labor:freiburg_2016_18.09.16_gst-assay_02_.xlsx">freiburg_2016_18.09.16_gst-assay_02_.xlsx</a>
+                 <p> All data and graphs were collected in an Excel file:<a href="https://static.igem.org/mediawiki/2016/0/05/T--Freiburg--LabJournal_g5_xlsx08.xlsx" class="media mediafile mf_xlsx" title="labor:freiburg_2016_18.09.16_gst-assay_02_.xlsx">freiburg_2016_18.09.16_gst-assay_02_.xlsx</a>
                      <br/>
                      <br/> </p>
              </div>
-             <!-- EDIT26 SECTION "GST-Assay 02" [16292-17526] -->
+             <!-- EDIT26 SECTION "GST-assay 02" [16292-17526] -->
              <h4 class="sectionedit27"><a name="gst_assay_03" id="gst_assay_03">GST – Assay 03</a></h4>
-             <div class="level3">
+             <div class="level3_1">
-                 <p> Repeated the assay of GST – Assay 02
+                 <p> Repeated the assay of GST – assay 01
-                     <br/> Pipetting scheme as in GST – Assay 02 (fig.26)
+                     <br/> Pipetting scheme as in GST – assay 01 (fig.26)
                      <br/>
                      <br/> </p>
@@ Line 756: / Line 757: @@
                  <ul>
                      <li class="level1">
-                         <div class="li"> Start the reaction with 90µl assay cocktail in each well</div>
+                         <div class="li"> Start the reaction with 90 µl master mix in each well</div>
                      </li>
                      <li class="level1">
@@ Line 767: / Line 768: @@
              <h5><a name="results2" id="results2">Results</a></h5>
              <div class="level4">
-                 <p> All graphs were collected in an excel file: <a href="https://static.igem.org/mediawiki/2016/1/14/T--Freiburg--LabJournal_g5_xlsx10.xlsx" class="media mediafile mf_xlsx" title="labor:freiburg_2016_gst_run3.xlsx">freiburg_2016_gst_run3.xlsx</a>
+                 <p> All graphs were collected in an Excel file: <a href="https://static.igem.org/mediawiki/2016/1/14/T--Freiburg--LabJournal_g5_xlsx10.xlsx" class="media mediafile mf_xlsx" title="labor:freiburg_2016_gst_run3.xlsx">freiburg_2016_gst_run3.xlsx</a>
                      <br/>
                      <br/> </p>
              </div>
-             <!-- EDIT27 SECTION "GST – Assay 03" [17527-17922] -->
+             <!-- EDIT27 SECTION "GST – assay 03" [17527-17922] -->
              <h4 class="sectionedit28"><a name="gst_assay_04" id="gst_assay_04">GST – Assay 04</a></h4>
-             <div class="level3">
+             <div class="level3_1">
-                 <p> Repeat the assay depending on the results of GST – Assay 03 (Freiburg 2016_gst_run3.xlsx). increased the measuring time up to 10min and pipetted on ice.
+                 <p> Repeat the assay depending on the results of GST – assay 03 (Freiburg 2016_gst_run3.xlsx). increased the measuring time up to 10 min and pipetted on ice.
                      <br/> Pipetted after scheme shown in fig.26.
                      <br/>
@@ Line 783: / Line 784: @@
                  <ul>
                      <li class="level1">
-                         <div class="li"> Start the reaction with 90µl assay cocktail in each well</div>
+                         <div class="li"> Start the reaction with 90 µl master mix in each well</div>
                      </li>
                      <li class="level1">
@@ Line 797: / Line 798: @@
              <h5><a name="results3" id="results3">Results</a></h5>
              <div class="level4">
-                 <p> All graphs were collected in an excel file: <a href="https://static.igem.org/mediawiki/2016/2/24/T--Freiburg--LabJournal_g5_xlsx12.xlsx" class="media mediafile mf_xlsx" title="labor:freiburg_2016_gst_run4.xlsx">freiburg_2016_gst_run4.xlsx</a>
+                 <p> All graphs were collected in an Excel file: <a href="https://static.igem.org/mediawiki/2016/2/24/T--Freiburg--LabJournal_g5_xlsx12.xlsx" class="media mediafile mf_xlsx" title="labor:freiburg_2016_gst_run4.xlsx">freiburg_2016_gst_run4.xlsx</a>
                      <br/>
                      <br/> </p>
              </div>
-             <!-- EDIT28 SECTION "GST – Assay 04" [17923-18467] -->
+             <!-- EDIT28 SECTION "GST – assay 04" [17923-18467] -->
              <h3 class="sectionedit29"><a name="section29082016" id="section29082016">29.08.2016</a></h3>
              <div class="level2"> </div>
              <!-- EDIT29 SECTION "29.08.2016" [18468-18490] -->
              <h4 class="sectionedit30"><a name="detergent_screening" id="detergent_screening">Detergent screening</a></h4>
-             <div class="level3">
+             <div class="level3_1">
                  <p> Scheme:
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:freiburg_2016_detergent_screening.png" class="media" title="labor:freiburg_2016_detergent_screening.png"><img src="https://static.igem.org/mediawiki/2016/e/ed/T--Freiburg--LabJournal_g5_27.png" class="media" alt="" /></a>
-                     <br/> fig.27: scheme of detergent screening for SDS, Tx100, Tween20, Shampoo1 and Shampoo2, as positive control was used a solution of 2% SDS, for negative control was used a wash solution of PBS without detergents.
+                     <br/> fig.27: scheme of detergent screening for SDS, Tx100, Tween20, Shampoo1 and Shampoo2, as positive control a solution of 2% SDS was used, as negative control a wash solution of PBS without detergents was used.
                      <br/>
                      <br/> </p>
@@ Line 818: / Line 819: @@
                  <ol>
                      <li class="level1">
-                         <div class="li"> Prepared GFP coating solution (20µg/ml)</div>
+                         <div class="li"> Prepared GFP coating solution (20 µg/ml)</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Coated the plate after the scheme (fig.27) with coating volume of 100µl</div>
+                         <div class="li"> Coated the plate after the scheme (fig.27) with coating volume of 100 µl</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Incubated for 2h at room temperature</div>
+                         <div class="li"> Incubated for 2 h at room temperature</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Washed the wells twice with 200µl PBS</div>
+                         <div class="li"> Washed the wells twice with 200 µl PBS</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Blocked wells with 200µl 5% nonfat dry milk in PBS</div>
+                         <div class="li"> Blocked wells with 200 µl 5% nonfat dry milk in PBS</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Incubated for 2h at room temperature</div>
+                         <div class="li"> Incubated for 2 h at room temperature</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Washed wells twice with 200µl PBS</div>
+                         <div class="li"> Washed wells twice with 200 µl PBS</div>
                      </li>
                  </ol>
@@ Line 846: / Line 847: @@
              <div class="level2">
                  <p> Preparation of detergents for screening: Prepared stock solutions of 20% and 40%
-                     <br/> - Prepared samples of 1ml for each the three cycles (except Block three there were only two cycles) in each block:
+                     <br/> - Prepared samples of 1 ml for each of the three cycles in each block (except block three there were only two cycles):
                      <br/> </p>
                  <ul>
                      <li class="level1">
-                         <div class="li"> Block 1: 2, 4, 6, 8% solutions of Tx100, Tween20, Shampoo1 (SH1) and Sampoo2 (SH2); for SDS used concentrations 1.5, 0.25, 1, 0.5 and 0.1% solutions</div>
+                         <div class="li"> block 1: 2, 4, 6, 8% solutions of Tx100, Tween20, Shampoo1 (SH1) and Shampoo2 (SH2); for SDS used concentrations 1.5, 0.25, 1, 0.5 and 0.1% solutions</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Block 2: 12, 14, 16,1 8% solutions of Tx100, Tween20, Shampoo1 (SH1) and Sampoo2 (SH2); for SDS used concentrations 1.5, 0.75, 1, 0.5 and 0.1% solutions</div>
+                         <div class="li"> block 2: 12, 14, 16,1 8% solutions of Tx100, Tween20, Shampoo1 (SH1) and Shampoo2 (SH2); for SDS used concentrations 1.5, 0.75, 1, 0.5 and 0.1% solutions</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Block 3: 32, 34,3 6, 38% solutions of Tx100, Tween20, Shampoo1 (SH1) and Sampoo2 (SH2); for SDS used concentrations 1.5, 1.25, 1, 0.5 and 0.1% solutions</div>
+                         <div class="li"> block 3: 32, 34,3 6, 38% solutions of Tx100, Tween20, Shampoo1 (SH1) and Shampoo2 (SH2); for SDS used concentrations 1.5, 1.25, 1, 0.5 and 0.1% solutions</div>
                      </li>
                  </ul>
@@ Line 864: / Line 865: @@
              <!-- EDIT31 SECTION "30.08.2016" [19165-19857] -->
              <h4 class="sectionedit32"><a name="washing" id="washing">Washing</a></h4>
-             <div class="level3">
+             <div class="level3_1">
                  <p> <strong>Block 1:</strong>
                      <br/> </p>
                  <ol>
                      <li class="level1">
-                         <div class="li">Washed with concentrations of 2, 4, 6, 8% solutions of Tx100, Tween20, Shampoo1 (SH1) and Sampoo2 (SH2); for SDS used concentrations 1.5, 0.25, 1, 0.5 and 0.1%, for negative control used PBS and for positive control 2% SDS. Wash solution volume took 200µl.</div>
+                         <div class="li">Washed with solutions with concentrations of 2, 4, 6, 8% of Tx100, Tween20, Shampoo1 (SH1) and Sampoo2 (SH2); for SDS used concentrations 1.5, 0.25, 1, 0.5 and 0.1%, for negative control used PBS and for positive control 2% SDS. Wash solution volume took 200 µl.</div>
                      </li>
                      <li class="level1">
@@ Line 881: / Line 882: @@
                      </li>
                      <li class="level1">
-                         <div class="li">repeated it for a total of 3 cycle</div>
+                         <div class="li">repeated it for a total of 3 cycles</div>
                      </li>
                  </ol>
                  <p>
-                     <br/> <strong>Block 2:</strong>
+                     <br/> <strong>block 2:</strong>
                      <br/> </p>
                  <ol>
                      <li class="level1">
-                         <div class="li"> Washed with concentrations of 12,14, 16, 18% solutions of Tx100, Tween20, Shampoo1 (SH1) and Sampoo2 (SH2); for SDS used concentrations 1.5, 0.75, 1, 0.5 and 0.1%, for negative control used PBS and for positive control 2% SDS. Wash solution volume took 200µl.</div>
+                         <div class="li"> Washed with solutions with concentrations of 12,14, 16, 18% of Tx100, Tween20, Shampoo1 (SH1) and Shampoo2 (SH2); for SDS used concentrations 1.5, 0.75, 1, 0.5 and 0.1%, for negative control used PBS and for positive control 2% SDS. Wash solution volume took 200 µl.</div>
                      </li>
                      <li class="level1">
@@ Line 901: / Line 902: @@
                      </li>
                      <li class="level1">
-                         <div class="li"> repeated it for a total of 3 cycle</div>
+                         <div class="li"> repeated it for a total of 3 cycles</div>
                      </li>
                  </ol>
                  <p>
-                     <br/> <strong>Block 3:</strong>
+                     <br/> <strong>block 3:</strong>
                      <br/> </p>
                  <ol>
                      <li class="level1">
-                         <div class="li"> Washed with concentrations of 32,34, 36, 38% solutions of Tx100, Tween20, Shampoo1 (SH1) and Sampoo2 (SH2); for SDS used concentrations 1.5, 1.25, 1, 0.5 and 0.1%, for negative control used PBS and for positive control 2% SDS. Wash solution volume took 200µl.</div>
+                         <div class="li"> Washed with solutions with concentrations of 32,34, 36, 38% of Tx100, Tween20, Shampoo1 (SH1) and Shampoo2 (SH2); for SDS used concentrations 1.5, 1.25, 1, 0.5 and 0.1%, for negative control used PBS and for positive control 2% SDS. Wash solution volume took 200 µl.</div>
                      </li>
                      <li class="level1">
@@ Line 929: / Line 930: @@
              <!-- EDIT32 SECTION "Washing" [19858-21200] -->
              <h4 class="sectionedit33"><a name="result6" id="result6">Result</a></h4>
-             <div class="level3">
+             <div class="level3_1">
-                 <p> Data and figures were collected in an excel file:<a href="https://static.igem.org/mediawiki/2016/d/d2/T--Freiburg--LabJournal_g5_xlsx13.xlsx" class="media mediafile mf_xlsx" title="labor:freiburg_2016_30.08.16_detergengt_screening.xlsx">freiburg_2016_30.08.16_detergengt_screening.xlsx</a>
+                 <p> Data and graphs were collected in an Excel file:<a href="https://static.igem.org/mediawiki/2016/d/d2/T--Freiburg--LabJournal_g5_xlsx13.xlsx" class="media mediafile mf_xlsx" title="labor:freiburg_2016_30.08.16_detergengt_screening.xlsx">freiburg_2016_30.08.16_detergengt_screening.xlsx</a>
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:freiburg_2016_overview_detergent_screening.png" class="media" title="labor:freiburg_2016_overview_detergent_screening.png"><img src="https://static.igem.org/mediawiki/2016/8/8e/T--Freiburg--LabJournal_g5_28.png" class="media" alt="" /></a>
@@ Line 942: / Line 943: @@
              <!-- EDIT34 SECTION "02.09.2016" [21458-21480] -->
              <h4 class="sectionedit35"><a name="washing_assay_03" id="washing_assay_03">Washing – Assay 03</a></h4>
-             <div class="level3">
+             <div class="level3_1">
                  <p> Plate name W5
                      <br/> coated the plate with GFP after coating protocol, used scheme from fig.29
@@ Line 952: / Line 953: @@
                      <br/>
                      <br/> stock solution 40% </p>
-                 <p> 12% | 300µl stock solution + 700µl PBS
+                 <p> 12% | 300 µl stock solution + 700 µl PBS
-                     <br/> 14% | 350µl stock solution + 650µl PBS
+                     <br/> 14% | 350 µl stock solution + 650 µl PBS
-                     <br/> 16% | 400µl stock solution + 600µl PBS
+                     <br/> 16% | 400 µl stock solution + 600 µl PBS
-                     <br/> 18% | 450µl stock solution + 550µl PBS
+                     <br/> 18% | 450 µl stock solution + 550 µl PBS
-                     <br/> 22% | 550µl stock solution + 450µl PBS
+                     <br/> 22% | 550 µl stock solution + 450 µl PBS
-                     <br/> 24% | 600µl stock solution + 400µl PBS
+                     <br/> 24% | 600 µl stock solution + 400 µl PBS
-                     <br/> 26% | 650µl stock solution + 350µl PBS
+                     <br/> 26% | 650 µl stock solution + 350 µl PBS
-                     <br/> 28% | 700µl stock solution + 300µl PBS
+                     <br/> 28% | 700 µl stock solution + 300 µl PBS
-                     <br/> 32% | 800µl stock solution + 200µl PBS
+                     <br/> 32% | 800 µl stock solution + 200 µl PBS
-                     <br/> 34% | 850µl stock solution + 150µl PBS
+                     <br/> 34% | 850 µl stock solution + 150 µl PBS
-                     <br/> 36% | 900µl stock solution + 100µl PBS
+                     <br/> 36% | 900 µl stock solution + 100 µl PBS
-                     <br/> 38% | 950µl stock solution + 50µl PBS
+                     <br/> 38% | 950 µl stock solution + 50 µl PBS
                      <br/>
                      <br/> </p>
@@ Line 969: / Line 970: @@
              <h5><a name="measured10" id="measured10">measured</a></h5>
              <div class="level4">
-                 <p> used setting for measuring the fluorescence form coated GFP and washed plate as shown in the protocol:
+                 <p> used setting for measuring the fluorescence from coated GFP and washed plate as shown in the protocol:
                      <br/> </p>
              </div>
              <h5><a name="result7" id="result7">Result</a></h5>
              <div class="level4">
-                 <p> Detailed figures collected in an excel file:
+                 <p> Detailed figures collected in an Excel file:
                      <br/>
                      <br/> Fig.30: overview of the wash efficiency of detergents: SDS: 1.25, 1, 0.75, 0.25%; Tween20: 22, 24, 26, 28, 32, 24, 26, 28%; SH1: 12, 14, 16 ,18, 22, 24, 26, 28%. For neg. contr. used 2% SDS, pos. contr. was washed with PBS without detergent.
@@ Line 980: / Line 981: @@
                      <br/> </p>
              </div>
-             <!-- EDIT35 SECTION "Washing – Assay 03" [21481-22885] -->
+             <!-- EDIT35 SECTION "Washing – assay 03" [21481-22885] -->
              <h4 class="sectionedit36"><a name="gst_assay_05" id="gst_assay_05">GST – Assay 05</a></h4>
-             <div class="level3">
+             <div class="level3_1">
-                 <p> repeated the assay depending on the results of GST – Assay 04 (excel file: Freiburg 2016gst_run4.xlsx.
+                 <p> repeated the assay depending on the results of GST – assay 04 (Excel file: Freiburg 2016gst_run4.xlsx.
                      <br/>
                      <br/> scheme:
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:freiburg_2016_gst_2_scheme.png" class="media" title="labor:freiburg_2016_gst_2_scheme.png"><img src="https://static.igem.org/mediawiki/2016/9/99/T--Freiburg--LabJournal_g5_26.png" class="media" alt="" /></a>
-                     <br/> fig.26: pipetting scheme of GST-amount (0.15U, 0.125U, 0.1U, 0.075U, 0.05U, 0.025U, 0.01U), white empty (Assay cocktail and PBS) in black Blank (Assay cocktail and PBS).
+                     <br/> fig.26: pipetting scheme of GST amounts (0.15 U, 0.125 U, 0.1 U, 0.075 U, 0.05 U, 0.025 U, 0.01 U), empty (master mix and PBS); blank (master mix and PBS).
                      <br/> </p>
                  <ol>
@@ Line 995: / Line 996: @@
                      </li>
                      <li class="level1">
-                         <div class="li"> incubated the plate before started in the fridge for 5min.</div>
+                         <div class="li"> incubated the plate before starting in the fridge for 5 min.</div>
                      </li>
                      <li class="level1">
@@ Line 1,006: / Line 1,007: @@
                  <ol>
                      <li class="level1">
-                         <div class="li"> started the reaction with 90µl assay cocktail in each well</div>
+                         <div class="li"> started the reaction with 90 µl master mix in each well</div>
                      </li>
                      <li class="level1">
@@ Line 1,030: / Line 1,031: @@
      <div class="color7">
          <div class="para_20">
-             <!-- EDIT36 SECTION "GST – Assay 05" [22886-23733] -->
+             <!-- EDIT36 SECTION "GST – assay 05" [22886-23733] -->
              <h3 class="sectionedit37"><a name="section03092016" id="section03092016">03.09.2016</a></h3>
              <div class="level2"> </div>
              <!-- EDIT37 SECTION "03.09.2016" [23734-23756] -->
-             <h4 class="sectionedit38"><a name="gst_assay_06" id="gst_assay_06">GST – Assay 06</a></h4>
+             <h4 class="sectionedit38"><a name="gst_assay_06" id="gst_assay_06">GST – assay 06</a></h4>
-             <div class="level3">
+             <div class="level3_1">
-                 <p> Preparation as like in GST – Assay 05. Took a little more time to start the measuring after mixing enzyme and assay cocktail
+                 <p> Preparation as in GST – assay 05. Took a little more time to start the measuring after mixing enzyme and master mix
                      <br/> Used pipetting scheme as shown in fig.26.
                      <br/>
@@ Line 1,049: / Line 1,050: @@
              <h5><a name="result9" id="result9">Result</a></h5>
              <div class="level4">
-                 <p> All data and graphs were collected in an excel file: <a href="https://static.igem.org/mediawiki/2016/a/a7/T--Freiburg--LabJournal_g5_xlsx17.xlsx" class="media mediafile mf_xlsx" title="labor:freiburg_2016_gst_run6.xlsx">freiburg_2016_gst_run6.xlsx</a>
+                 <p> All data and graphs were collected in an Excel file: <a href="https://static.igem.org/mediawiki/2016/a/a7/T--Freiburg--LabJournal_g5_xlsx17.xlsx" class="media mediafile mf_xlsx" title="labor:freiburg_2016_gst_run6.xlsx">freiburg_2016_gst_run6.xlsx</a>
                      <br/>
                      <br/> </p>
              </div>
-             <!-- EDIT38 SECTION "GST – Assay 06" [23757-24177] -->
+             <!-- EDIT38 SECTION "GST – assay 06" [23757-24177] -->
              <h4 class="sectionedit39"><a name="gst_assay_07" id="gst_assay_07">GST – Assay 07</a></h4>
-             <div class="level3">
+             <div class="level3_1">
-                 <p> Repeated like as in assay 05, but all was pipetted on ice. Used pipetting scheme as shown in fig.26.
+                 <p> Repeated like as in assay 05, but everything was pipetted on ice. Used pipetting scheme as shown in fig.26.
                      <br/>
                      <br/> </p>
@@ Line 1,062: / Line 1,063: @@
              <h5><a name="measured12" id="measured12">Measured</a></h5>
              <div class="level4">
-                 <p> Start the reaction with 90µl assay cocktail from low amount of GST concentration to high, for reducing the time delay the assay cocktail was piped in directly next to the plate reader
+                 <p> Start the reaction with 90 µl master mix from low GST concentration to high, for reducing the time delay the master mix was pipetted directly next to the plate reader
                      <br/>
                      <br/> </p>
@@ Line 1,068: / Line 1,069: @@
              <h5><a name="result10" id="result10">Result</a></h5>
              <div class="level4">
-                 <p> All data and Graphs were collected in an excel file: <a href="https://static.igem.org/mediawiki/2016/c/cb/T--Freiburg--LabJournal_g5_xlsx18.xlsx" class="media mediafile mf_xlsx" title="labor:freibur_2016_gst_run7.xlsx">freibur_2016_gst_run7.xlsx</a>
+                 <p> All data and graphs were collected in an Excel file: <a href="https://static.igem.org/mediawiki/2016/c/cb/T--Freiburg--LabJournal_g5_xlsx18.xlsx" class="media mediafile mf_xlsx" title="labor:freibur_2016_gst_run7.xlsx">freibur_2016_gst_run7.xlsx</a>
                      <br/>
                      <br/> </p>
              </div>
-             <!-- EDIT39 SECTION "GST – Assay 07" [24178-24629] -->
+             <!-- EDIT39 SECTION "GST – assay 07" [24178-24629] -->
              <h4 class="sectionedit40"><a name="gst_assay_08" id="gst_assay_08">GST – Assay 08</a></h4>
-             <div class="level3"> </div>
+             <div class="level3_1"> </div>
              <h5><a name="preparation3" id="preparation3">Preparation</a></h5>
              <div class="level4">
-                 <p> Preparations like as in the other Assays before (GST – Assay 07) after pipetted and incubate for 5min in the fridge put the plate on a shaker.
+                 <p> Preparations like in the other assays before (GST – assay 07) after pipetting and incubating for 5 min in the fridge, put the plate on a shaker.
                      <br/>
                      <br/> </p>
@@ Line 1,083: / Line 1,084: @@
              <h5><a name="measuring2" id="measuring2">Measuring</a></h5>
              <div class="level4">
-                 <p> started the reaction with 90 µl Assay cocktail while shaking at 300rpm and temperature (temperature of the block) of 12°C.
+                 <p> started the reaction with 90 µl master mix while shaking at 300rpm and shaker temperature of 12°C.
-                     <br/> Put it then directly into the reader.
+                     <br/> Put it then into the reader.
                      <br/> protocol: <a href="https://static.igem.org/mediawiki/2016/a/a4/T--Freiburg--LabJournal_g5_xlsx19.xlsx" class="media mediafile mf_xlsx" title="labor:protocol_gst_run_8.xlsx">protocol_gst_run_8.xlsx</a>
                      <br/>
@@ Line 1,091: / Line 1,092: @@
              <h5><a name="result11" id="result11">Result</a></h5>
              <div class="level4">
-                 <p> graphs were collected in an excel file: <a href="https://static.igem.org/mediawiki/2016/8/84/T--Freiburg--LabJournal_g5_xlsx20.xlsx" class="media mediafile mf_xlsx" title="labor:freiburg_2016_gst_run8.xlsx">freiburg_2016_gst_run8.xlsx</a>
+                 <p> graphs were collected in an Excel file: <a href="https://static.igem.org/mediawiki/2016/8/84/T--Freiburg--LabJournal_g5_xlsx20.xlsx" class="media mediafile mf_xlsx" title="labor:freiburg_2016_gst_run8.xlsx">freiburg_2016_gst_run8.xlsx</a>
                      <br/>
                      <br/> </p>
              </div>
-             <!-- EDIT40 SECTION "GST – Assay 08" [24630-25166] -->
+             <!-- EDIT40 SECTION "GST – assay 08" [24630-25166] -->
              <h4 class="sectionedit41"><a name="gst-assay_09" id="gst-assay_09">GST-Assay 09</a></h4>
-             <div class="level3"> </div>
+             <div class="level3_1"> </div>
              <h5><a name="preparation4" id="preparation4">Preparation</a></h5>
              <div class="level4">
-                 <p> * as in GST-Assay 8 * increased measuring time up to 20min in the measuring settings
+                 <p> * as in GST-assay 8 * increased measuring time up to 20 min in the measuring settings
                      <br/>
                      <br/> </p>
@@ Line 1,106: / Line 1,107: @@
              <h5><a name="measuring3" id="measuring3">Measuring</a></h5>
              <div class="level4">
-                 <p> Started the reaction like in GST-Assay 8, used settings as shown in the protocol: <a href="https://static.igem.org/mediawiki/2016/4/4c/T--Freiburg--LabJournal_g5_xlsx21.xlsx" class="media mediafile mf_xlsx" title="labor:protocol_gst_run_9.xlsx">protocol_gst_run_9.xlsx</a>
+                 <p> Started the reaction like in GST-assay 8, used settings as shown in the protocol: <a href="https://static.igem.org/mediawiki/2016/4/4c/T--Freiburg--LabJournal_g5_xlsx21.xlsx" class="media mediafile mf_xlsx" title="labor:protocol_gst_run_9.xlsx">protocol_gst_run_9.xlsx</a>
                      <br/>
                      <br/> </p>
@@ Line 1,112: / Line 1,113: @@
              <h5><a name="result12" id="result12">Result</a></h5>
              <div class="level4">
-                 <p> All data and graphs were in an excel file: <a href="https://static.igem.org/mediawiki/2016/5/52/T--Freiburg--LabJournal_g5_xlsx22.xlsx" class="media mediafile mf_xlsx" title="labor:freiburg_2016_gst_run9.xlsx">freiburg_2016_gst_run9.xlsx</a>
+                 <p> All data and graphs were in an Excel file: <a href="https://static.igem.org/mediawiki/2016/5/52/T--Freiburg--LabJournal_g5_xlsx22.xlsx" class="media mediafile mf_xlsx" title="labor:freiburg_2016_gst_run9.xlsx">freiburg_2016_gst_run9.xlsx</a>
                      <br/>
                      <br/> </p>
              </div>
-             <!-- EDIT41 SECTION "GST-Assay 09" [25167-25549] -->
+             <!-- EDIT41 SECTION "GST-assay 09" [25167-25549] -->
              <h3 class="sectionedit42"><a name="section12092016" id="section12092016">12.09.2016</a></h3>
              <div class="level2"> </div>
              <!-- EDIT42 SECTION "12.09.2016" [25550-25572] -->
              <h4 class="sectionedit43"><a name="gst_assay_10" id="gst_assay_10">GST – Assay 10</a></h4>
-             <div class="level3"> </div>
+             <div class="level3_1"> </div>
-             <h5><a name="perpetrated" id="perpetrated">Perpetrated</a></h5>
+             <h5><a name="Preparation" id="Preparation">Preparation</a></h5>
              <div class="level4"> </div>
              <h6><a name="samples_of_gst" id="samples_of_gst">samples of GST:</a></h6>
@@ Line 1,128: / Line 1,129: @@
                  <ul>
                      <li class="level1">
-                         <div class="li"> 0.15U (90µl of 5U/ml stock)</div>
+                         <div class="li"> 0.15 U (90 µl of 5 U/ml stock)</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> 0.075U (15µl of 5U/ml stock + 75µl PBS)</div>
+                         <div class="li"> 0.075 U (15µl of 5 U/ml stock + 75 µl PBS)</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> 0.025U (5µl of 5U/ml stock + 85µl PBS)</div>
+                         <div class="li"> 0.025 U (5 µl of 5 U/ml stock + 85 µl PBS)</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> 0,005U (3µl of 5U/ml stock + 87µl PBS)</div>
+                         <div class="li"> 0,005 U (3 µl of 5 U/ml stock + 87 µl PBS)</div>
                      </li>
                  </ul>
                  <p>
                      <br/>
-                     <br/> Assay cocktail: 15µl CDNB + 15µl GSH + 1470µl PBS.
+                     <br/> master mix: 15 µl CDNB + 15 µl GSH + 1470 µl PBS.
-                     <br/> Used 30µl enzyme solution and 90µl Assay cocktail per well, for Blank and Empty used 30µl PBS and Assay cocktail.
+                     <br/> Used 30 µl enzyme solution and 90 µl master mix per well, for blank and empty used 30 µl PBS and master mix.
                      <br/>
                      <br/> Scheme:
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:gst_scheme_12.09.16_01.png" class="media" title="labor:gst_scheme_12.09.16_01.png"><img src="https://static.igem.org/mediawiki/2016/a/a8/T--Freiburg--LabJournal_g5_31.png" class="media" alt="" /></a>
-                     <br/> fig.31: Pipetting scheme for GST – Assay wit 0.15, 0.075, 0.025 and 0.005 Units of GST and Assay cocktail, for wells empty and blank used instead of GST solutions PBS and Assay cocktail.
+                     <br/> fig.31: Pipetting scheme for GST – assay with 0.15, 0.075, 0.025 and 0.005 Units of GST and master mix, for wells empty and blank used instead of GST solutions PBS and master mix.
                      <br/>
-                     <br/> Assay cocktail wasn’t enough for all wells, concentration of 0.15U without assay cocktail.
+                     <br/> master mix wasn’t enough for all wells, concentration of 0.15 U without master mix.
                      <br/>
                      <br/> </p>
@@ Line 1,162: / Line 1,163: @@
              <h5><a name="result13" id="result13">Result</a></h5>
              <div class="level4">
-                 <p> Data and Graphs were collected in an excel file:
+                 <p> Data and graphs were collected in an Excel file:
                      <br/>
                      <br/> </p>
              </div>
-             <!-- EDIT43 SECTION "GST – Assay 10" [25573-26477] -->
+             <!-- EDIT43 SECTION "GST – assay 10" [25573-26477] -->
-             <h4 class="sectionedit44"><a name="gst_assay_11" id="gst_assay_11">GST – Assay 11</a></h4>
+             <h4 class="sectionedit44"><a name="gst_assay_11" id="gst_assay_11">GST – assay 11</a></h4>
-             <div class="level3">
+             <div class="level3_1">
-                 <p> Repeated GST – Assay 10 with more assay cocktail, scheme fig.31
+                 <p> Repeated GST – assay 10 with more master mix, scheme fig.31
-                     <br/> 1,7ml: 17µl CDNB + 17µl GSH + 1666µl PBS
+                     <br/> 1,7 ml: 17 µl CDNB + 17 µl GSH + 1666 µl PBS
                      <br/>
                      <br/> </p>
@@ Line 1,182: / Line 1,183: @@
              <h5><a name="result14" id="result14">Result</a></h5>
              <div class="level4">
-                 <p> Data and Graphs were collected in an excel file:
+                 <p> Data and graphs were collected in an Excel file:
                      <br/>
                      <br/> </p>
              </div>
-             <!-- EDIT44 SECTION "GST – Assay 11" [26478-26755] -->
+             <!-- EDIT44 SECTION "GST – assay 11" [26478-26755] -->
-             <h4 class="sectionedit45"><a name="gst_assay_12" id="gst_assay_12">GST – Assay 12</a></h4>
+             <h4 class="sectionedit45"><a name="gst_assay_12" id="gst_assay_12">GST – assay 12</a></h4>
-             <div class="level3">
+             <div class="level3_1">
-                 <p> Repeated measurement of GST – Assay 11 with different measurement settings.
+                 <p> Repeated measurement of GST – assay 11 with different measurement settings.
-                     <br/> Used scheme the same scheme fig.31
+                     <br/> Used the same scheme fig.31
                      <br/>
                      <br/> </p>
@@ Line 1,202: / Line 1,203: @@
              <h5><a name="result15" id="result15">Result</a></h5>
              <div class="level4">
-                 <p> Data and Graphs were collected in an excel file:
+                 <p> Data and graphs were collected in an Excel file:
                      <br/>
                      <br/> </p>
              </div>
-             <!-- EDIT45 SECTION "GST – Assay 12" [26756-27036] -->
+             <!-- EDIT45 SECTION "GST – assay 12" [26756-27036] -->
              <h3 class="sectionedit46"><a name="section14092016" id="section14092016">14.09.2016</a></h3>
              <div class="level2"> </div>
              <!-- EDIT46 SECTION "14.09.2016" [27037-27059] -->
              <h4 class="sectionedit47"><a name="shampoowashing_with_nanobody" id="shampoowashing_with_nanobody">Shampoo: washing with nanobody</a></h4>
-             <div class="level3"> </div>
+             <div class="level3_1"> </div>
              <h5><a name="prepared" id="prepared">Prepared</a></h5>
              <div class="level4">
                  <p> Coated plate after the scheme
-                     <br/> Prepared nanobody solution: Concentration nanobody stock solution 0.48mg/ml, set up dilution of 1.8ng/µl
+                     <br/> Prepared nanobody solution: Concentration nanobody stock solution 0.48 mg/ml, set up dilution of 1.8 ng/µl
                      <br/> Scheme:
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:washed_with_naonobody_1.png" class="media" title="labor:washed_with_naonobody_1.png"><img src="https://static.igem.org/mediawiki/2016/e/eb/T--Freiburg--LabJournal_g5_32.png" class="media" alt="" /></a>
-                     <br/> fig.32: washing scheme for washing with nanobody and detergents. As control were used: GFP washed with PBS (negative control 1), washed with nanobody in PBS (negative control 2), 2%SDS (positive control). As reference were empty wells not coated and unwashed, same as the blank. Used detergent samples: SDS: 1, 0.75, 0.5, 0.25% solutions with nanobody, Tween20: 22, 24, 26,28% solutions with nanobody.
+                     <br/> fig.32: washing scheme for washing with nanobody and detergents. As control were used: GFP washed with PBS (negative control 1), washed with nanobody in PBS (negative control 2), 2%SDS (positive control). As reference empty wells were not coated and unwashed, same as the blank. Used detergent samples: SDS: 1, 0.75, 0.5, 0.25% solutions with nanobody, Tween20: 22, 24, 26, 28% solutions with nanobody.
                      <br/>
                      <br/> </p>
@@ Line 1,230: / Line 1,231: @@
                      </li>
                      <li class="level1">
-                         <div class="li"> Washed with 1µl nanobody and 200µl concentrations of detergents: SDS: 1.5, 1, 0.5, 0.25%, Tween20: 22, 24, 26, 28%, SH1: 12, 14, 16, 18%</div>
+                         <div class="li"> Washed with 1 µl nanobody and 200 µl concentrations of detergents: SDS: 1.5, 1, 0.5, 0.25%, Tween20: 22, 24, 26, 28%, SH1: 12, 14, 16, 18%</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Incubated it for 5min</div>
+                         <div class="li"> Incubated it for 5 min</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> Shook it for 5min at 300rpm</div>
+                         <div class="li"> Shook it for 5 min at 300 rpm</div>
                      </li>
                      <li class="level1">
@@ Line 1,253: / Line 1,254: @@
              <h5><a name="result16" id="result16">Result</a></h5>
              <div class="level4">
-                 <p> Detailed column charts were collected in an excel file: <a href="https://static.igem.org/mediawiki/2016/d/dd/T--Freiburg--LabJournal_g5_xlsx23.xlsx" class="media mediafile mf_xlsx" title="labor:washing_with_nanobody_eff..xlsx">washing_with_nanobody_eff..xlsx</a>
+                 <p> Detailed column charts were collected in an Excel file: <a href="https://static.igem.org/mediawiki/2016/d/dd/T--Freiburg--LabJournal_g5_xlsx23.xlsx" class="media mediafile mf_xlsx" title="labor:washing_with_nanobody_eff..xlsx">washing_with_nanobody_eff..xlsx</a>
                      <br/>
                      <br/>
@@ Line 1,266: / Line 1,267: @@
              <!-- EDIT48 SECTION "14.09.2016" [28707-28729] -->
              <h4 class="sectionedit49"><a name="gst_assay_13" id="gst_assay_13">GST – Assay 13</a></h4>
-             <div class="level3">
+             <div class="level3_1">
                  <p> Scheme:
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:gst_scheme_12.09.16_01.png" class="media" title="labor:gst_scheme_12.09.16_01.png"><img src="https://static.igem.org/mediawiki/2016/a/a8/T--Freiburg--LabJournal_g5_31.png" class="media" alt="" /></a>
-                     <br/> fig.34: pipetting scheme of GST – Assay, used GST amount of: 0.0001, 0.00001, 0.000001, 0.0000001 Units, for empty and blank was used PBS instead of GST.
+                     <br/> fig.34: pipetting scheme of GST – assay, used GST amount of: 0.0001, 0.00001, 0.000001, 0.0000001 Units, for empty and blank was used PBS instead of GST.
                      <br/>
                      <br/> </p>
@@ Line 1,286: / Line 1,287: @@
                      <br/> </p>
              </div>
-             <!-- EDIT49 SECTION "GST – Assay 13" [28730-29215] -->
+             <!-- EDIT49 SECTION "GST – assay 13" [28730-29215] -->
              <h3 class="sectionedit50"><a name="section16092016" id="section16092016">16.09.2016</a></h3>
              <div class="level2"> </div>
              <!-- EDIT50 SECTION "16.09.2016" [29216-29238] -->
              <h4 class="sectionedit51"><a name="shampoo" id="shampoo">Shampoo</a></h4>
-             <div class="level3">
+             <div class="level3_1">
                  <p> Platename: WN2
                      <br/> Scheme:
@@ Line 1,302: / Line 1,303: @@
                  <ol>
                      <li class="level1">
-                         <div class="li"> coated plate with GFP (coating concentration 20µg/ml)</div>
+                         <div class="li"> coated plate with GFP (coating concentration 20 µg/ml)</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> dilute 1µl nanobody stock solution (0.48mg/ml) in prepared samples 1ml detergents: SDS: 1, 0.75, 0.5, 0.25, Tween20: 22, 24, 26, 28%, SH1: 12, 14, 16, 18%</div>
+                         <div class="li"> dilute 1 µl nanobody stock solution (0.48 mg/ml) in prepared samples 1 ml detergents: SDS: 1, 0.75, 0.5, 0.25, Tween20: 22, 24, 26, 28%, SH1: 12, 14, 16, 18%</div>
                      </li>
                  </ol>
@@ Line 1,313: / Line 1,314: @@
              <!-- EDIT51 SECTION "Shampoo" [29239-29973] -->
              <h4 class="sectionedit52"><a name="wash-assay" id="wash-assay">Wash-assay</a></h4>
-             <div class="level3">
+             <div class="level3_1">
                  <ol>
                      <li class="level1">
@@ Line 1,319: / Line 1,320: @@
                      </li>
                      <li class="level1">
-                         <div class="li"> Washed with 200µl detergents: SDS: 1.5, 1, 0.5, 0.25% (Row A and B with nanobody, row C and D without nanobody), Tween20: 22, 24, 26, 28% (Row A and B with nanobody, row C and D without nanobody), SH1: 12, 14, 16, 18% (Row E and F with nanobody, row G and H without nanobody)</div>
+                         <div class="li"> Washed with 200 µl detergents: SDS: 1.5, 1, 0.5, 0.25% (row A and B with nanobody, row C and D without nanobody), Tween20: 22, 24, 26, 28% (row A and B with nanobody, row C and D without nanobody), SH1: 12, 14, 16, 18% (row E and F with nanobody, row G and H without nanobody)</div>
                      </li>
                      <li class="level1">
-                         <div class="li">Incubated it for 5min</div>
+                         <div class="li">Incubated it for 5 min</div>
                      </li>
                      <li class="level1">
-                         <div class="li">Shook it for 5min at 300rpm</div>
+                         <div class="li">Shook it for 5 min at 300 rpm</div>
                      </li>
                      <li class="level1">
@@ Line 1,346: / Line 1,347: @@
              <h5><a name="result18" id="result18">Result</a></h5>
              <div class="level4">
-                 <p> Detailed column charts were collected in an excel file:<a href="https://static.igem.org/mediawiki/2016/0/03/T--Freiburg--LabJournal_g5_xlsx26.xlsx" class="media mediafile mf_xlsx" title="labor:freiburg_2016_16.09.16_washing_nanobody_02_gfp_master.xlsx">freiburg_2016_16.09.16_washing_nanobody_02_gfp_master.xlsx</a>
+                 <p> Detailed column charts were collected in an Excel file:<a href="https://static.igem.org/mediawiki/2016/0/03/T--Freiburg--LabJournal_g5_xlsx26.xlsx" class="media mediafile mf_xlsx" title="labor:freiburg_2016_16.09.16_washing_nanobody_02_gfp_master.xlsx">freiburg_2016_16.09.16_washing_nanobody_02_gfp_master.xlsx</a>
                      <br/>
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:freiburg_2016_overview_of_the_wash_efficiency_between_with_and_without_nanobody.png" class="media" title="labor:freiburg_2016_overview_of_the_wash_efficiency_between_with_and_without_nanobody.png"><img src="https://static.igem.org/mediawiki/2016/c/c6/T--Freiburg--LabJournal_g5_36.png" class="media" alt="" /></a>
-                     <br/> Fig.36: Overview of the wash efficiency between with and without nanobody for: SDS, SDS + nanobody, Tween20, Twenn20 + nanobody, SH1, SH1 + nanobody.
+                     <br/> Fig.36: Overview of the wash efficiency with and without nanobody for: SDS, SDS + nanobody, Tween20, Twenn20 + nanobody, SH1, SH1 + nanobody.
                      <br/>
                      <br/> </p>
@@ Line 1,359: / Line 1,360: @@
              <!-- EDIT53 SECTION "17.09.2016" [30980-31002] -->
              <h4 class="sectionedit54"><a name="gst_assay_14" id="gst_assay_14">GST – Assay 14</a></h4>
-             <div class="level3">
+             <div class="level3_1">
-                 <p> Repeat the GST – Assay 13 with the same scheme (fig34).
+                 <p> Repeat the GST – assay 13 with the same scheme (fig34).
                      <br/> Concentration of samples:
                      <br/> </p>
                  <ul>
                      <li class="level1">
-                         <div class="li"> 0,0001U (3µl 0,1U/ml stock + 87µl PBS)</div>
+                         <div class="li"> 0,000 1U (3 µl 0,1 U/ml stock + 87 µl PBS)</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> 0,00001U (3µl 0,01U/ml stock + 87 PBS)</div>
+                         <div class="li"> 0,00001 U (3 µl 0,01 U/ml stock + 87µl PBS)</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> 0,000001U (3µl 0,001U/ml stock + 87 PBS)</div>
+                         <div class="li"> 0,000001 U (3 µl 0,001 U/ml stock + 87µl PBS)</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> 0,0000001U (3µl 0,0001U/ml stock + 87 PBS)</div>
+                         <div class="li"> 0,0000001 U (3 µl 0,0001 U/ml stock + 87µl PBS)</div>
                      </li>
                  </ul>
                  <p>
-                     <br/> Assay cocktail: 15µl CDNB + 15µl GSH + 1470µl PBS.
+                     <br/> master mix: 15 µl CDNB + 15 µl GSH + 1470 µl PBS.
-                     <br/> Pipetted the GST after the scheme (30µl per well), used for empty wells and blank instead of GST solution PBS.
+                     <br/> Pipetted the GST after the scheme (30 µl per well), used for empty wells and blank instead of GST solution PBS.
                      <br/>
                      <br/> </p>
@@ Line 1,385: / Line 1,386: @@
              <h5><a name="measured17" id="measured17">Measured</a></h5>
              <div class="level4">
-                 <p> Start the reaction with 90µ assay cocktail from low concentration to high concentration, pipetted directly next to the plate reader to recuse the time delay.
+                 <p> Start the reaction with 90 µl mastr mix from low concentration to high concentration, pipetted next to the plate reader to reduce the time delay.
-                     <br/> Measured the absorbents with settings as described in the measurement protocol: <a href="https://static.igem.org/mediawiki/2016/c/c2/T--Freiburg--LabJournal_g5_xlsx48.xlsx" class="media mediafile mf_xlsx" title="labor:17.09.16_gst-assay_01_settings.xlsx">17.09.16_gst-assay_01_settings.xlsx</a>
+                     <br/> Measured the absorbance with settings as described in the measurement protocol: <a href="https://static.igem.org/mediawiki/2016/c/c2/T--Freiburg--LabJournal_g5_xlsx48.xlsx" class="media mediafile mf_xlsx" title="labor:17.09.16_gst-assay_01_settings.xlsx">17.09.16_gst-assay_01_settings.xlsx</a>
                      <br/>
                      <br/> </p>
              </div>
-             <!-- EDIT54 SECTION "GST – Assay 14" [31003-31787] -->
+             <!-- EDIT54 SECTION "GST – assay 14" [31003-31787] -->
-             <h4 class="sectionedit55"><a name="gst_assay_15" id="gst_assay_15">GST – Assay 15</a></h4>
+             <h4 class="sectionedit55"><a name="gst_assay_15" id="gst_assay_15">GST – assay 15</a></h4>
-             <div class="level3">
+             <div class="level3_1">
-                 <p> Repeated the GST – Assay 14
+                 <p> Repeated the GST – assay 14
                      <br/> settings: <a href="https://static.igem.org/mediawiki/2016/e/e5/T--Freiburg--LabJournal_g5_xlsx27.xlsx" class="media mediafile mf_xlsx" title="labor:17.09.16_gst-assay_02_settings.xlsx">17.09.16_gst-assay_02_settings.xlsx</a>
                      <br/>
                      <br/> </p>
              </div>
-             <!-- EDIT55 SECTION "GST – Assay 15" [31788-31910] -->
+             <!-- EDIT55 SECTION "GST – assay 15" [31788-31910] -->
-             <h4 class="sectionedit56"><a name="gst_assay_16" id="gst_assay_16">GST – Assay 16</a></h4>
+             <h4 class="sectionedit56"><a name="gst_assay_16" id="gst_assay_16">GST – assay 16</a></h4>
-             <div class="level3">
+             <div class="level3_1">
-                 <p> Changed setting for the measurement: shook in plate reader before each cycle at 500rpm 1s.
+                 <p> Changed setting for the measurement: shook in plate reader before each cycle at 500 rpm 1 s.
                      <br/>
                      <br/> Scheme:
                      <br/>
                      <a href="https://static.igem.org/mediawiki/2016/e/eb/T--Freiburg--LabJournal_g5_xlsx28.xlsx" class="media" title="labor:gst_scheme_17.09.16_02.png"><img src="https://static.igem.org/mediawiki/2016/1/10/T--Freiburg--LabJournal_g5_37.png" class="media" alt="" /></a>
-                     <br/> fig.37: GST – Assay pipetting scheme for GST concentrations of 0.005 and 0.0000001 Units. Empty as negative control with PBS instead of GST, blank as the same conditions like empty.
+                     <br/> fig.37: GST – assay pipetting scheme for GST concentrations of 0.005 and 0.0000001 Units. Empty as negative control with PBS instead of GST, blank with the same conditions like empty.
                      <br/>
                      <br/> prepared </p>
                  <ul>
                      <li class="level1">
-                         <div class="li"> samples of 0.005 and 0.0000001U</div>
+                         <div class="li"> samples of 0.005 and 0.0000001 U</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> assay – cocktail (14µl CDNB + 14µl GSH + 1372µl PBS)</div>
+                         <div class="li"> master mix (14 µl CDNB + 14 µl GSH + 1372 µl PBS)</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> pipetted GST-samples after the scheme and PBS for the controls (30µl)</div>
+                         <div class="li"> pipetted GST samples after the scheme and PBS for the controls (30 µl)</div>
                      </li>
                  </ul>
@@ Line 1,425: / Line 1,426: @@
              <h5><a name="measurement" id="measurement">Measurement</a></h5>
              <div class="level4">
-                 <p> start the reaction with 170µl Assay cocktail. Used new setting as in the reader protocol described: <a href="https://static.igem.org/mediawiki/2016/8/8d/T--Freiburg--LabJournal_g5_xlsx29.xlsx" class="media mediafile mf_xlsx" title="labor:17.09.16_gst-assay_03_settings.xlsx">17.09.16_gst-assay_03_settings.xlsx</a>
+                 <p> start the reaction with 170 µl master mix. Used new setting as in the reader protocol described: <a href="https://static.igem.org/mediawiki/2016/8/8d/T--Freiburg--LabJournal_g5_xlsx29.xlsx" class="media mediafile mf_xlsx" title="labor:17.09.16_gst-assay_03_settings.xlsx">17.09.16_gst-assay_03_settings.xlsx</a>
                      <br/>
                      <br/> </p>
              </div>
-             <!-- EDIT56 SECTION "GST – Assay 16" [31911-32639] -->
+             <!-- EDIT56 SECTION "GST – assay 16" [31911-32639] -->
              <h4 class="sectionedit57"><a name="result19" id="result19">Result</a></h4>
-             <div class="level3">
+             <div class="level3_1">
-                 <p> All data and graphs of GST - Assay 14 - 16 were collected in an excel file: <a href="https://static.igem.org/mediawiki/2016/7/78/T--Freiburg--LabJournal_g5_xlsx30.xlsx" class="media mediafile mf_xlsx" title="labor:freiburg_2016_17.09.16_gst-assay_master.xlsx">freiburg_2016_17.09.16_gst-assay_master.xlsx</a>
+                 <p> All data and graphs of GST - assay 14 - 16 were collected in an Excel file: <a href="https://static.igem.org/mediawiki/2016/7/78/T--Freiburg--LabJournal_g5_xlsx30.xlsx" class="media mediafile mf_xlsx" title="labor:freiburg_2016_17.09.16_gst-assay_master.xlsx">freiburg_2016_17.09.16_gst-assay_master.xlsx</a>
                      <br/>
                      <br/> </p>
@@ Line 1,447: / Line 1,448: @@
              <!-- EDIT58 SECTION "18.09.2016" [32795-32817] -->
              <h4 class="sectionedit59"><a name="gst_assay_17" id="gst_assay_17">GST – Assay 17</a></h4>
-             <div class="level3">
+             <div class="level3_1">
                  <p> Doubled the amount of substrate
                      <br/> Scheme:
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:gst_scheme_18.09.16.png" class="media" title="labor:gst_scheme_18.09.16.png"><img src="https://static.igem.org/mediawiki/2016/5/56/T--Freiburg--LabJournal_g5_38.png" class="media" alt="" /></a>
-                     <br/> Fig.38: Pipetting scheme for GST – Assay with 0.1, 0.025, 0.005 and 0.001 Units (U), empty wells and blank measured with PBS instead of GST as control.
+                     <br/> Fig.38: Pipetting scheme for GST – assay with 0.1, 0.025, 0.005 and 0.001 Units (U), empty wells and blank measured with PBS instead of GST as control.
                      <br/>
                      <br/> </p>
@@ Line 1,458: / Line 1,459: @@
              <h5><a name="prepared2" id="prepared2">Prepared</a></h5>
              <div class="level4">
-                 <p> GST – samples:
+                 <p> GST samples:
                      <br/> </p>
                  <ul>
                      <li class="level1">
-                         <div class="li"> 0.1U (60µl of 5U/ml stock + 30µl PBS)</div>
+                         <div class="li"> 0.1 U (60 µl of 5 U/ml stock + 30 µl PBS)</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> 0.025U (5µl of 5U/ml stock + 87µl PBS)</div>
+                         <div class="li"> 0.025 U (5 µl of 5 U/ml stock + 87 µl PBS)</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> 0.005U (3µl of 5U/ml stock + 85µl PBS)</div>
+                         <div class="li"> 0.005 U (3 µl of 5 U/ml stock + 85 µl PBS)</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> 0,001U (3µl of 1U/ml stock + 87µl PBS)</div>
+                         <div class="li"> 0,001 U (3 µl of 1 U/ml stock + 87 µl PBS)</div>
                      </li>
                  </ul>
-                 <p> Assay cocktail: 30µl CDNB + 30µl GSH + 1440µl PBS.
+                 <p> master mix: 30 µl CDNB + 30 µl GSH + 1440 µl PBS.
-                     <br/> Pipet GST – samples and PBS after the scheme (fig.38).
+                     <br/> Pipet GST samples and PBS after the scheme (fig.38).
                      <br/> </p>
              </div>
              <h5><a name="measured18" id="measured18">Measured</a></h5>
              <div class="level4">
-                 <p> Start the reaction with 90µl assay cocktail
+                 <p> Start the reaction with 90 µl master mix
                      <br/> Start measuring with settings which are described in the protocol: <a href="https://static.igem.org/mediawiki/2016/2/2f/T--Freiburg--LabJournal_g5_xlsx31.xlsx" class="media mediafile mf_xlsx" title="labor:18.09.16_gst-assay_01_settings.xlsx">18.09.16_gst-assay_01_settings.xlsx</a>
                      <br/>
@@ Line 1,487: / Line 1,488: @@
              <h5><a name="result20" id="result20">Result</a></h5>
              <div class="level4">
-                 <p> Data and graphs were collected in an excel file:<a href="https://static.igem.org/mediawiki/2016/1/13/T--Freiburg--LabJournal_g5_xlsx32.xlsx" class="media mediafile mf_xlsx" title="labor:18.09.16_gst-assay_01.xlsx">18.09.16_gst-assay_01.xlsx</a>
+                 <p> Data and graphs were collected in an Excel file:<a href="https://static.igem.org/mediawiki/2016/1/13/T--Freiburg--LabJournal_g5_xlsx32.xlsx" class="media mediafile mf_xlsx" title="labor:18.09.16_gst-assay_01.xlsx">18.09.16_gst-assay_01.xlsx</a>
                      <br/>
                      <br/> </p>
              </div>
-             <!-- EDIT59 SECTION "GST – Assay 17" [32818-33715] -->
+             <!-- EDIT59 SECTION "GST – assay 17" [32818-33715] -->
-             <h4 class="sectionedit60"><a name="gst_assay_18" id="gst_assay_18">GST – Assay 18</a></h4>
+             <h4 class="sectionedit60"><a name="gst_assay_18" id="gst_assay_18">GST – assay 18</a></h4>
-             <div class="level3">
+             <div class="level3_1">
                  <p> Scheme:
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:gst_scheme_18.09.16_02.png" class="media" title="labor:gst_scheme_18.09.16_02.png"><img src="https://static.igem.org/mediawiki/2016/8/8a/T--Freiburg--LabJournal_g5_39.png" class="media" alt="" /></a>
-                     <br/> fig.39: Pipetting scheme for GST – Assay with 0.1, 0.05 Units (U), empty wells and blank measured with PBS instead of GST as control, grey wells are not used.
+                     <br/> fig.39: Pipetting scheme for GST assay with 0.1, 0.05 Units (U), empty wells and blank measured with PBS instead of GST as control, grey wells were not used.
                      <br/>
                      <br/> Increased the measuring time up to 33 minutes.
@@ Line 1,505: / Line 1,506: @@
              <h5><a name="prepared3" id="prepared3">Prepared</a></h5>
              <div class="level4">
-                 <p> GST – Samples:
+                 <p> GST Samples:
                      <br/> </p>
                  <ul>
                      <li class="level1">
-                         <div class="li"> 0,1U</div>
+                         <div class="li"> 0,1 U</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> 0,05U (30µl of 5U/ml stock + 60µl PBS)</div>
+                         <div class="li"> 0,05 U (30 µl of 5 U/ml stock + 60 µl PBS)</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> 0,025U
+                         <div class="li"> 0,025 U
                              <br/> </div>
                      </li>
                  </ul>
-                 <p> Assay cocktail
+                 <p> master mix
                      <br/>
                      <br/> </p>
@@ Line 1,525: / Line 1,526: @@
              <h5><a name="measurement1" id="measurement1">Measurement</a></h5>
              <div class="level4">
-                 <p> Start reactions with 90µl assay cocktail Used setting as decried in the measurement protocol of the reader: <a href="https://static.igem.org/mediawiki/2016/0/02/T--Freiburg--LabJournal_g5_xlsx33.xlsx" class="media mediafile mf_xlsx" title="labor:18.09.16_gst-assay_02_settings.xlsx">18.09.16_gst-assay_02_settings.xlsx</a>
+                 <p> Start reactions with 90 µl master mix. Used setting as described in the measurement protocol of the reader: <a href="https://static.igem.org/mediawiki/2016/0/02/T--Freiburg--LabJournal_g5_xlsx33.xlsx" class="media mediafile mf_xlsx" title="labor:18.09.16_gst-assay_02_settings.xlsx">18.09.16_gst-assay_02_settings.xlsx</a>
                      <br/>
                      <br/> </p>
@@ Line 1,531: / Line 1,532: @@
              <h5><a name="result21" id="result21">Result</a></h5>
              <div class="level4">
-                 <p> Data and graphs were collected in an excel file: <a href="https://static.igem.org/mediawiki/2016/2/20/T--Freiburg--LabJournal_g5_xlsx34.xlsx" class="media mediafile mf_xlsx" title="labor:freiburg_2016_18.09.16_gst-assay_02_-1.xlsx">freiburg_2016_18.09.16_gst-assay_02_-1.xlsx</a>
+                 <p> Data and graphs were collected in an Excel file: <a href="https://static.igem.org/mediawiki/2016/2/20/T--Freiburg--LabJournal_g5_xlsx34.xlsx" class="media mediafile mf_xlsx" title="labor:freiburg_2016_18.09.16_gst-assay_02_-1.xlsx">freiburg_2016_18.09.16_gst-assay_02_-1.xlsx</a>
                      <br/>
                      <br/> </p>
              </div>
-             <!-- EDIT60 SECTION "GST – Assay 18" [33716-34451] -->
+             <!-- EDIT60 SECTION "GST – assay 18" [33716-34451] -->
              <h3 class="sectionedit61"><a name="section19092016" id="section19092016">19.09.2016</a></h3>
              <div class="level2"> </div>
              <!-- EDIT61 SECTION "19.09.2016" [34452-34474] -->
-             <h4 class="sectionedit62"><a name="gst_assay_19" id="gst_assay_19">GST – Assay 19</a></h4>
+             <h4 class="sectionedit62"><a name="gst_assay_19" id="gst_assay_19">GST – assay 19</a></h4>
-             <div class="level3">
+             <div class="level3_1">
                  <p> Scheme:
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:gst_scheme_19.09.16_01.png" class="media" title="labor:gst_scheme_19.09.16_01.png"><img src="https://static.igem.org/mediawiki/2016/3/3f/T--Freiburg--LabJournal_g5_40.png" class="media" alt="" /></a>
-                     <br/> fig.40: Pipetting scheme for GST – Assay with 0.1, 0.05 Units (U), empty wells and blank measured with PBS instead of GST as control, grey wells are not used.
+                     <br/> fig.40: Pipetting scheme for GST – assay with 0.1, 0.05 Units (U), empty wells and blank measured with PBS instead of GST as control, grey wells were not used.
                      <br/>
                      <br/> </p>
@@ Line 1,554: / Line 1,555: @@
                  <ul>
                      <li class="level1">
-                         <div class="li"> 0,1U (80µl 5U/ml stock + 40µl PBS)</div>
+                         <div class="li"> 0,1 U (80 µl 5 U/ml stock + 40 µl PBS)</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> 0,05U (40µl 5U/ml stock + 80µl PBS)</div>
+                         <div class="li"> 0,05 U (40 µl 5 U/ml stock + 80 µl PBS)</div>
                      </li>
                  </ul>
-                 <p> Assay cocktail: 18µl CDNB + 18µl GSH + 1164µl PBS
+                 <p> master mix: 18 µl CDNB + 18 µl GSH + 1164 µl PBS
-                     <br/> Changed shook time before the first cycle in the setting to 200rpm for 1s
+                     <br/> Changed shaking time before the first cycle in the setting to 200 rpm for 1s
-                     <br/> Piped the samples after the scheme in fig.42
+                     <br/> Pipetted the samples after the scheme in fig.42
                      <br/>
                      <br/> </p>
@@ Line 1,568: / Line 1,569: @@
              <h5><a name="measurement2" id="measurement2">Measurement</a></h5>
              <div class="level4">
-                 <p> Start the reactions with 90µl PBS and start measurement with the setting as described in the plate reader protocol: <a href="https://static.igem.org/mediawiki/2016/4/42/T--Freiburg--LabJournal_g5_xlsx35.xlsx" class="media mediafile mf_xlsx" title="labor:19.09.16_gst-assay_01_settings.xlsx">19.09.16_gst-assay_01_settings.xlsx</a>
+                 <p> Start the reactions with 90 µl master mix and start measurement with the setting as described in the plate reader protocol: <a href="https://static.igem.org/mediawiki/2016/4/42/T--Freiburg--LabJournal_g5_xlsx35.xlsx" class="media mediafile mf_xlsx" title="labor:19.09.16_gst-assay_01_settings.xlsx">19.09.16_gst-assay_01_settings.xlsx</a>
                      <br/>
                      <br/> </p>
              </div>
-             <!-- EDIT62 SECTION "GST – Assay 19" [34475-35219] -->
+             <!-- EDIT62 SECTION "GST – assay 19" [34475-35219] -->
-             <h4 class="sectionedit63"><a name="gst_assay_20" id="gst_assay_20">GST – Assay 20</a></h4>
+             <h4 class="sectionedit63"><a name="gst_assay_20" id="gst_assay_20">GST – assay 20</a></h4>
-             <div class="level3">
+             <div class="level3_1">
-                 <p> Repeat the GST – Assay 19 in addition with one well of enzyme without assay cocktail
+                 <p> Repeat the GST – assay 19 in addition with one well of enzyme without master mix
                      <br/> Scheme:
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:gst_scheme_19.09.16_02.png" class="media" title="labor:gst_scheme_19.09.16_02.png"><img src="https://static.igem.org/mediawiki/2016/1/1d/T--Freiburg--LabJournal_g5_41.png" class="media" alt="" /></a>
-                     <br/> Fig.41: Pipetting scheme for GST – Assay with 0.1, 0.05 Units (U), empty wells and blank measured with PBS instead of GST as control, En well which contains only enzyme without assay cocktail, grey wells are not used.
+                     <br/> Fig.41: Pipetting scheme for GST – assay with 0.1, 0.05 Units (U), empty wells and blank measured with PBS instead of GST as control, En = well which contains only enzyme without assay cocktail, grey wells were not used.
                      <br/>
                      <br/> </p>
@@ Line 1,589: / Line 1,590: @@
                  <ul>
                      <li class="level1">
-                         <div class="li"> 0,1U (80µl 5U/ml stock + 40µl PBS)</div>
+                         <div class="li"> 0,1 U (80 µl 5 U/ml stock + 40 µl PBS)</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> 0,05U (40µl 5U/ml stock + 80µl PBS)</div>
+                         <div class="li"> 0,05 U (40 µl 5 U/ml stock + 80 µl PBS)</div>
                      </li>
                  </ul>
                  <p>
-                     <br/> Assay cocktail: 18µl CDNB + 18µl GSH + 1164µl PBS
+                     <br/> master mix: 18 µl CDNB + 18 µl GSH + 1164 µl PBS
-                     <br/> For En (fig.41) used 30µl of 1U GST solution
+                     <br/> For En (fig.41) used 30 µl of 1 U GST solution
-                     <br/> Piped the samples after the scheme in fig.41
+                     <br/> Pipetted the samples after the scheme in fig.41
                      <br/>
                      <br/> </p>
@@ Line 1,604: / Line 1,605: @@
              <h5><a name="measurement3" id="measurement3">Measurement</a></h5>
              <div class="level4">
-                 <p> Start the reactions with 90µl PBS and start measuremen.
+                 <p> Start the reactions with 90 µl master mix and start measurement
                      <br/>
                      <br/> </p>
@@ Line 1,610: / Line 1,611: @@
              <h5><a name="result22" id="result22">Result</a></h5>
              <div class="level4">
-                 <p> All data and graphs of GST - Assay 19 and 20 were collected in an excel file: <a href="/igem2016/lib/exe/fetch.php?media=labor:freiburg_2016_19.09.16_gst-assay_master-1.xlsx" class="media mediafile mf_xlsx" title="labor:freiburg_2016_19.09.16_gst-assay_master-1.xlsx">freiburg_2016_19.09.16_gst-assay_master-1.xlsx</a>
+                 <p> All data and graphs of GST - assay 19 and 20 were collected in an Excel file: <a href="https://2016.igem.org/File:T--Freiburg--LabJournal_g5_xlsx36.xlsx" class="media mediafile mf_xlsx" title="labor:freiburg_2016_19.09.16_gst-assay_master-1.xlsx">freiburg_2016_19.09.16_gst-assay_master-1.xlsx</a>
                      <br/>
                      <br/> </p>
              </div>
-             <!-- EDIT63 SECTION "GST – Assay 20" [35220-36135] -->
+             <!-- EDIT63 SECTION "GST – assay 20" [35220-36135] -->
              <h3 class="sectionedit64"><a name="section23092016" id="section23092016">23.09.2016</a></h3>
              <div class="level2"> </div>
              <!-- EDIT64 SECTION "23.09.2016" [36136-36158] -->
-             <h4 class="sectionedit65"><a name="gst_assay_21" id="gst_assay_21">GST – Assay 21</a></h4>
+             <h4 class="sectionedit65"><a name="gst_assay_21" id="gst_assay_21">GST – assay 21</a></h4>
-             <div class="level3">
+             <div class="level3_1">
                  <p> Scheme:
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:gst_scheme_23.09.16_01.png" class="media" title="labor:gst_scheme_23.09.16_01.png"><img src="https://static.igem.org/mediawiki/2016/b/b5/T--Freiburg--LabJournal_g5_42.png" class="media" alt="" /></a>
-                     <br/> Fig.42: Pipetting scheme for GST – Assay with 0.1, empty wells and blank measured with PBS instead of GST as control, grey wells are not used.
+                     <br/> Fig.42: Pipetting scheme for GST – assay with 0.1 U, empty wells and blank measured with PBS instead of GST as control, grey wells were not used.
                      <br/>
                      <br/> </p>
@@ Line 1,629: / Line 1,630: @@
              <h5><a name="prepared6" id="prepared6">Prepared</a></h5>
              <div class="level4">
-                 <p> Sample of 0.1U and assay cocktail: 70µl CDNB + 70µl GSH + 560µl PBS, increased amount of substrates.
+                 <p> Sample of 0.1 U and master mix: 70 µl CDNB + 70 µl GSH + 560 µl PBS, increased amount of substrates.
                      <br/>
                      <br/> </p>
@@ Line 1,635: / Line 1,636: @@
              <h5><a name="measurement4" id="measurement4">Measurement</a></h5>
              <div class="level4">
-                 <p> Start the reaction with 90µ assay cocktail.
+                 <p> Start the reaction with 90 µl master mix.
                      <br/> Start the measurement wit settings as before, which are described also in the reader protocol: <a href="https://static.igem.org/mediawiki/2016/6/62/T--Freiburg--LabJournal_g5_xlsx36.xlsx" class="media mediafile mf_xlsx" title="labor:23.09.16_gst-assay_01_settings.xlsx">23.09.16_gst-assay_01_settings.xlsx</a>
                      <br/>
                      <br/> </p>
              </div>
-             <h5><a name="gst_assay_22" id="gst_assay_22">GST – Assay 22</a></h5>
+             <h5><a name="gst_assay_22" id="gst_assay_22">GST – assay 22</a></h5>
              <div class="level4">
-                 <p> Reduced amount of substrates from 70µl to 50µl and increased volume from 90µ to 110µl used blank of GST – Assay 21 (scheme fig.42)
+                 <p> Reduced amount of substrates from 70 µl to 50 µl and increased volume from 90 µl to 110 µl used blank of GST – assay 21 (scheme fig.42)
                      <br/> Scheme:
                      <br/>
                      <a href="/igem2016/lib/exe/fetch.php?media=labor:gst_scheme_23.09.16_02.png" class="media" title="labor:gst_scheme_23.09.16_02.png"><img src="https://static.igem.org/mediawiki/2016/9/9b/T--Freiburg--LabJournal_g5_43.png" class="media" alt="" /></a>
-                     <br/> Fig.43: Pipetting scheme for GST – Assay 22. 0.1Units GST for sample wells and as referents Empty which contains PBS and assay cocktail.
+                     <br/> Fig.43: Pipetting scheme for GST – assay 22. 0.1 Units GST for sample wells and as reference empty which contains PBS and master mix.
                      <br/>
                      <br/> </p>
@@ Line 1,654: / Line 1,655: @@
                  <ul>
                      <li class="level1">
-                         <div class="li"> Prepared 0.1U samples of GST and assay cocktail: 50µl CDNB + 50µl GSH + 900µl PBS</div>
+                         <div class="li"> Prepared 0.1 U samples of GST and master mix: 50 µl CDNB + 50 µl GSH + 900 µl PBS</div>
                      </li>
                      <li class="level1">
-                         <div class="li"> pipetted 30µl GST and PBS after the scheme (fig.43)</div>
+                         <div class="li"> pipetted 30 µl GST and PBS after the scheme (fig.43)</div>
                      </li>
                  </ul>
@@ Line 1,665: / Line 1,666: @@
              <h5><a name="measurement5" id="measurement5">Measurement</a></h5>
              <div class="level4">
-                 <p> Start the reactions with 110µl assay cocktail
+                 <p> Start the reactions with 110 µl master mix
                      <br/> started measurement with setting which are described in the plate reader protocol:<a href="https://static.igem.org/mediawiki/2016/c/c7/T--Freiburg--LabJournal_g5_xlsx38.xlsx" class="media mediafile mf_xlsx" title="labor:23.09.16_gst-assay_02_settings.xlsx">23.09.16_gst-assay_02_settings.xlsx</a>
                      <br/>
-                     <br/> Result: All data and graphs of GST - Assay 22 and 23 were collected in an excel file: <a href="https://static.igem.org/mediawiki/2016/3/39/T--Freiburg--LabJournal_g5_xlsx39.xlsx" class="media mediafile mf_xlsx" title="labor:freiburg_23.09.16_gst-assay_master-1.xlsx">freiburg_23.09.16_gst-assay_master-1.xlsx</a>
+                     <br/> Result: All data and graphs of GST - assay 22 and 23 were collected in an Excel file: <a href="https://static.igem.org/mediawiki/2016/3/39/T--Freiburg--LabJournal_g5_xlsx39.xlsx" class="media mediafile mf_xlsx" title="labor:freiburg_23.09.16_gst-assay_master-1.xlsx">freiburg_23.09.16_gst-assay_master-1.xlsx</a>
                      <br/>
                      <br/> </p>
@@ Line 1,676: / Line 1,677: @@
 </span></div>
              </div>
-             <!-- EDIT65 SECTION "GST – Assay 21" [36159-] -->
+             <!-- EDIT65 SECTION "GST – assay 21" [36159-] -->
          </div>
          <!-- para20 -->
@@ Line 1,689: / Line 1,690: @@
                  <p> with <em>E.coli</em> cell lysate
                      <br/> neg. control: DH5α
-                     <br/> pos. control: 0,05U GST
+                     <br/> pos. control: 0,05 U GST
                      <br/> constructs 131, 134 &amp; 138 (concentrations: 0,05 0,075 0,1 [µg/µl] protein per well)
                      <br/>
@@ Line 1,702: / Line 1,703: @@
              <!-- EDIT67 SECTION "29.09.2016" [37886-37908] -->
              <h4 class="sectionedit68"><a name="gst_assay1" id="gst_assay1">GST assay</a></h4>
-             <div class="level3">
+             <div class="level3_1">
-                 <p> concentrations: 0,1U 0,08U 0,06U 0,04U
+                 <p> concentrations: 0,1 U 0,08 U 0,06 U 0,04 U
-                     <br/> 0,08U: 48µl 5U/ml stock + 60µl PBS
+                     <br/> 0,08 U: 48 µl 5 U/ml stock + 60 µl PBS
-                     <br/> 0,06U: 36µl stock + 72µl PBS
+                     <br/> 0,06 U: 36 µl stock + 72 µl PBS
-                     <br/> 0,04U: 24µl stock + 96µl PBS
+                     <br/> 0,04 U: 24 µl stock + 96 µl PBS
-                     <br/> 1,7ml assaycocktail: 85µl CDNB + 85µl GSH + 1530µl PBS
+                     <br/> 1,7 ml master mix: 85 µl CDNB + 85 µl GSH + 1530 µl PBS
-                     <br/> 40µl enzyme solution + 100µl assaycocktail per well
+                     <br/> 40 µl enzyme solution + 100 µl master mix per well
                      <br/>
                      <br/> Didn&#039;t work, maybe pipetting error.
@@ Line 1,720: / Line 1,721: @@
              <!-- EDIT69 SECTION "30.09.2016" [38254-38276] -->
              <h4 class="sectionedit70"><a name="gst_assay2" id="gst_assay2">GST assay</a></h4>
-             <div class="level3">
+             <div class="level3_1">
                  <p> <em>E.coli</em> lysate
                      <br/>
                      <br/> construct 134
-                     <br/> 15µl per well: 30µl lysate + 150µl PBS → 69µg proteins per well
+                     <br/> 15 µl per well: 30 µl lysate + 150 µl PBS → 69 µg proteins per well
-                     <br/> 30µl per well: 60µl lysate + 120µl PBS → 138µg proteins per well
+                     <br/> 30 µl per well: 60 µl lysate + 120 µl PBS → 138 µg proteins per well
-                     <br/> 60µl per well: 120µl lysate + 60µl PBS → 276µg proteins per well
+                     <br/> 60 µl per well: 120 µl lysate + 60 µl PBS → 276 µg proteins per well
-                     <br/> 90µl per well: 180µl lysate → 414µg proteins per well
+                     <br/> 90 µl per well: 180 µl lysate → 414 µg proteins per well
-                     <br/> 60µl + GST per well: 120µl lysate + 0,05U (20µl 5U/ml stock + 40µl PBS)
+                     <br/> 60 µl + GST per well: 120 µl lysate + 0,05 U (20 µl 5 U/ml stock + 40 µl PBS)
                      <br/>
                      <br/> DH5α
-                     <br/> 30µl per well: 60µl lysate + 120µl PBS → 57µg proteins per well
+                     <br/> 30 µl per well: 60 µl lysate + 120 µl PBS → 57 µg proteins per well
                      <br/>
-                     <br/> 0,1U
+                     <br/> 0,1 U
-                     <br/> 40µl 5U/ml stock + 140µl PBS
+                     <br/> 40 µl 5 U/ml stock + 140 µl PBS
                      <br/>
-                     <br/> 1,5ml assaycocktail: 75µl CDNB + 75µl GSH + 1350µl PBS
+                     <br/> 1,5ml master mix: 75 µl CDNB + 75 µl GSH + 1350 µl PBS
                      <br/>
-                     <br/> 90µl sample + 90µl assaycocktail per well
+                     <br/> 90 µl sample + 90 µl master mix per well
                      <br/>
                      <br/>
@@ Line 1,747: / Line 1,748: @@
              <!-- EDIT70 SECTION "GST assay" [38277-38989] -->
              <h4 class="sectionedit71"><a name="gst_assay3" id="gst_assay3">GST assay</a></h4>
-             <div class="level3">
+             <div class="level3_1">
                  <p> <em>E.coli</em> lysate
                      <br/> constructs 131 &amp; 138
                      <br/>
                      <br/> 131
-                     <br/> 28µl → 148,4µg proteins
+                     <br/> 28 µl → 148,4 µg proteins
-                     <br/> 52µl → 275,6µg proteins
+                     <br/> 52 µl → 275,6 µg proteins
-                     <br/> 75µl → 397,5µg proteins
+                     <br/> 75 µl → 397,5 µg proteins
                      <br/>
                      <br/> 138
-                     <br/> 25µl → 147,5µg proteins
+                     <br/> 25 µl → 147,8 µg proteins
-                     <br/> 47µl → 277,3µg proteins
+                     <br/> 47 µl → 277,3 µg proteins
-                     <br/> 68µl → 401,2µg proteins
+                     <br/> 68 µl → 401,2 µg proteins
                      <br/>
                      <br/> DH5α
-                     <br/> 79µl → 150,1µg proteins
+                     <br/> 79 µl → 150,1 µg proteins
                      <br/>
                      <br/> DH5α+
-                     <br/> 79µl + 0,05U (10µl stock + 1µl PBS)
+                     <br/> 79 µl + 0,05 U (10 µl stock + 1 µl PBS)
                      <br/>
-                     <br/> 0,05U
+                     <br/> 0,05 U
                      <br/>
-                     <br/> 1,9ml assaycocktail: 95µl CDNB + 95µl GSH + 1710µl PBS
+                     <br/> 1,9 ml master mix: 95 µl CDNB + 95 µl GSH + 1710 µl PBS
                      <br/>
-                     <br/> 90µl sample + 90µl assaycocktail
+                     <br/> 90 µl sample + 90 µl master mix
                      <br/>
                      <br/>
@@ Line 1,793: / Line 1,794: @@
              <!-- EDIT72 SECTION "02.10.2016" [39522-39544] -->
              <h4 class="sectionedit73"><a name="gst_assay4" id="gst_assay4">GST assay</a></h4>
-             <div class="level3">
+             <div class="level3_1">
                  <p> GST-spores
                      <br/> constructs: Z+GST &amp; WT+GST
                      <br/> concentrations: 50 million, 25 million, 10 million, 5 million spores
-                     <br/> stock 100 million spores in 500µl
+                     <br/> stock 100 million spores in 500 µl
-                     <br/> 50 million: 250µl stock per well
+                     <br/> 50 million: 250 µl stock per well
-                     <br/> 25 million: 125µl stock per well + 125µl PBS
+                     <br/> 25 million: 125 µl stock per well + 125 µl PBS
-                     <br/> 10 million: 50µl stock per well + 200µl PBS
+                     <br/> 10 million: 50 µl stock per well + 200 µl PBS
-                     <br/> 5 million: 25µl stock per well + 225µl PBS
+                     <br/> 5 million: 25 µl stock per well + 225 µl PBS
                      <br/>
-                     <br/> WT as control: 250µl per well
+                     <br/> WT as control: 250 µl per well
                      <br/>
-                     <br/> 0,05U GST
+                     <br/> 0,05 U GST
                      <br/>
-                     <br/> 1,4ml assaycocktail: 70µl CDNB + 70µl GSH + 1260µl PBS
+                     <br/> 1,4 ml master mix: 70 µl CDNB + 70 µl GSH + 1260 µl PBS
                      <br/>
-                     <br/> 250µl sample + 90µl assaycocktail per well
+                     <br/> 250 µl sample + 90 µl master mix per well
                      <br/>
                      <br/> done again with the CotZ + GST construct and the CotZ construct purified
@@ Line 1,827: / Line 1,828: @@
              <!-- EDIT74 SECTION "03.10.16" [40271-40292] -->
              <h4 class="sectionedit75"><a name="gst_assay5" id="gst_assay5">GST assay</a></h4>
-             <div class="level3">
+             <div class="level3_1">
-                 <p> measured <em>E.coli</em> lysates again with about 150µg, 275µg and 400µg of proteins
+                 <p> measured <em>E.coli</em> lysates again with about 150 µg, 275 µg and 400 µg of proteins
                      <br/>
                      <br/>
@@ Line 1,835: / Line 1,836: @@
              <h4><a name="results6" id="results6">Results</a></h4>
              <div class="level4">
-                 <p> The GST-spores showed no difference in activity compared to the wild type.
+                 <p> The GST spores showed no difference in activity compared to the wild type.
                      <br/>
                      <br/>
@@ Line 1,842: / Line 1,843: @@
              <!-- EDIT75 SECTION "GST assay" [40293-40503] -->
              <h4 class="sectionedit76"><a name="gst_assay6" id="gst_assay6">GST assay</a></h4>
-             <div class="level3">
+             <div class="level3_1">
                  <p> measured the WT + GST spores purified
-                     <br/> concentrations: 50million, 25million, 10million, 5million spores
+                     <br/> concentrations: 50 million, 25 million, 10 million, 5 million spores
-                     <br/> controls: pos. 0,05U GST &amp; neg. untransformed WT spores
+                     <br/> controls: pos. 0,05 U GST &amp; neg. untransformed WT spores
                      <br/>
                      <br/>
@@ Line 1,861: / Line 1,862: @@
              <!-- EDIT77 SECTION "04.10.16" [40747-40767] -->
              <h4 class="sectionedit78"><a name="gst-assay" id="gst-assay">GST-assay</a></h4>
-             <div class="level3">
+             <div class="level3_1">
-                 <p> measured assaycocktail with freshly prepared GSH in ethanol and freshly prepared GSH in demineralized water
+                 <p> measured master mix with freshly prepared GSH in ethanol and freshly prepared GSH in demineralized water
-                     <br/> 400µl assaycocktail: 20µl CDNB + 20µl GSH + 360µl PBS
+                     <br/> 400 µl master mix: 20 µl CDNB + 20 µl GSH + 360 µl PBS
                      <br/>
                      <br/>
@@ Line 1,886: / Line 1,887: @@
              <!-- EDIT79 SECTION "05.10.16" [41045-41065] -->
              <h4 class="sectionedit80"><a name="gst-assay1" id="gst-assay1">GST-assay</a></h4>
-             <div class="level3">
+             <div class="level3_1">
-                 <p> measured 0,1U; 0,08U; 0,06U; 0,04U
+                 <p> measured 0,1 U; 0,08 U; 0,06 U; 0,04 U
-                     <br/> 3ml assaycocktail: 150µl CDNB + 150µl GSH + 2700µl PBS
+                     <br/> 3 ml master mix: 150 µl CDNB + 150 µl GSH + 2700 µl PBS
                      <br/>
                      <br/> </p>
@@ Line 1,905: / Line 1,906: @@
              <!-- EDIT81 SECTION "06.10.16" [41343-41363] -->
              <h4 class="sectionedit82"><a name="gst-assay2" id="gst-assay2">GST-assay</a></h4>
-             <div class="level3">
+             <div class="level3_1">
                  <p> measured <em>E.coli</em> lysates
-                     <br/> DH5α: 6,8µg/µl proteins
+                     <br/> DH5α: 6,8 µg/µl proteins
-                     <br/> 138: 6,8µg/µl
+                     <br/> 138: 6,8 µg/µl
-                     <br/> 134: 6,3µg/µl
+                     <br/> 134: 6,3 µg/µl
                      <br/>
-                     <br/> concentrations: ~400µg; ~500µg; ~600µg per well
+                     <br/> concentrations: ~400 µg; ~500 µg; ~600 µg per well
                      <br/>
-                     <br/> controls: 0,05U, Empty(PBS+assaycocktail)
+                     <br/> controls: 0,05 U, empty (PBS+master mix)
                      <br/>
                      <br/>
@@ Line 1,920: / Line 1,921: @@
                      <br/>
                      <br/> <a href="https://static.igem.org/mediawiki/2016/9/98/T--Freiburg--LabJournal_g5_xlsx43.xlsx" class="media mediafile mf_xlsx" title="labor:06.10.16_gst-assay_lysates_settings.xlsx">06.10.16_gst-assay_lysates_settings.xlsx</a>
-                    <br/> <a href="https://static.igem.org/mediawiki/2016/5/5a/T--Freiburg--LabJournal_g5_xlsx44.xlsx" class="media mediafile mf_xlsx" title="labor:06.10.16_10.10.16_gst-assay_lysates_master.xlsx">06.10.16_10.10.16_gst-assay_lysates_master.xlsx</a>
                      <br/>
                      <br/>
-                     <br/> </p>
+                     <p> results see below 10.10.16
+                        <br/>
+                        <br/> </p>
              </div>
              <!-- EDIT82 SECTION "GST-assay" [41364-41750] -->
              <h4 class="sectionedit83"><a name="gst-assay3" id="gst-assay3">GST-assay</a></h4>
-             <div class="level3">
+             <div class="level3_1">
                  <p> measured CotZ-GST knock-out spores and WT spores
-                     <br/> 50million - 5 million spores
+                     <br/> 50 million - 5 million spores
                      <br/> washed unpurified spores 3 times with PBS
                      <br/>
                      <br/> controls:
-                     <br/> pos. 0,05U GST (4*10µl 5U/ml stock + 960µl PBS)
+                     <br/> pos. 0,05 U GST (4*10 µl 5 U/ml stock + 960 µl PBS)
-                     <br/> neg. empty (250µl PBS + 90µl assaycocktail)
+                     <br/> neg. empty (250 µl PBS + 90 µl master mix)
-                     <br/> untrans. WT (50-5million)
+                     <br/> untrans. WT (50-5 million)
                      <br/>
-                     <br/> 3ml assaycocktail: 150µl CDNB + 150µl GSH + 2700µl PBS
+                     <br/> 3 ml master mix: 150 µl CDNB + 150 µl GSH + 2700 µl PBS
                      <br/>
-                     <br/> 250µl sample + 90µl assaycocktail
+                     <br/> 250 µl sample + 90 µl master mix
                      <br/>
                      <br/>
@@ Line 1,950: / Line 1,952: @@
                      <br/>
                      <br/> </p>
+                <h2 class="sectionedit86"><a name="section081016" id="section081016">08.10.16</a></h2>
+                <div class="level2"> </div>
+                <!-- EDIT86 SECTION "08.10.16" [42226-42246] -->
+                <h3 class="sectionedit87"><a name="shampoo1" id="shampoo1">Shampoo</a></h3>
+                <div class="level3_1">
+                    <p> tested: aGFPnano-CotZ(159Z) spores </p>
+                </div>
+                <h4><a name="plate_preparation" id="plate_preparation">Plate preparation</a></h4>
+                <div class="level4">
+                    <p> coated 68 wells with GFP, coating concentration GFP 20 µg/ml and coating volume 100 µl
+                        <br/> incubated 2 h at room temerature
+                        <br/> washed twice with 200 µl PBS
+                        <br/> blocked with 200 µl 5% nonfat dry milk in PBS
+                        <br/> incubated for 2 h at roomtemperatur
+                        <br/> washed twice with PBS
+                        <br/> measured the fluorescence of coated GFP
+                        <br/>
+                        <br/> </p>
+                </div>
+                <h4><a name="washing1" id="washing1">Washing</a></h4>
+                <div class="level4">
+                    <p> used 200 µl washing of solutions:
+                        <br/> </p>
+                    <ul>
+                        <li class="level1">
+                            <div class="li"> SDS: 1%, 0.75%, 0.5%, 0.25% (with spores, nanobody and only SDS)</div>
+                        </li>
+                        <li class="level1">
+                            <div class="li"> Tween20: 22%, 24%, 26%, 28% (with spores, nanobody and only Tween20)</div>
+                        </li>
+                        <li class="level1">
+                            <div class="li"> SH1: 12%, 14%, 16%, 18% (with spores, nanobody and only SH1)</div>
+                        </li>
+                        <li class="level1">
+                            <div class="li"> Controls: nanobody in PBS, WT - spore in PBS, construct spore (159Z) in PBS, 2% SDS &amp; PBS </div>
+                        </li>
+                    </ul>
+                    <p>
+                        <br/> incubated wash solutions on the plate for 5 min
+                        <br/> shook for 5 min at 300 rpm
+                        <br/> washed twice with PBS
+                        <br/> measured the emission of remaining GFP
+                        <br/>
+                        <br/>
+                        <br/> </p>
+                </div>
+                <h4><a name="results12" id="results12">Results</a></h4>
+                <div class="level4">
+                    <p> <a href="https://static.igem.org/mediawiki/2016/9/98/T--Freiburg--deliveryjournal4.xlsx" title="labor:bba_k2114002_agfpnano_ha_g4s_cotz_in_δcotz_.xlsx">bba_k2114002_agfpnano_ha_g4s_cotz_in_δcotz_.xlsx</a>
+                        <br/>
+                        <br/>
+                        <br/> </p>
+                </div>
              </div>
              <!-- EDIT83 SECTION "GST-assay" [41751-42206] -->
@@ Line 1,956: / Line 2,011: @@
              <!-- EDIT84 SECTION "09.10.16" [42207-42227] -->
              <h4 class="sectionedit85"><a name="gst-assay4" id="gst-assay4">GST-assay</a></h4>
-             <div class="level3">
+             <div class="level3_1">
                  <p> with GST-CotZ(153)-, CotG-GST(156)-, GST-CotG(167)- &amp; CgeA-GST(168)-construct spores.
-                     <br/> 50million - 5million spores
+                     <br/> 50 million - 5 million spores
                      <br/>
                      <br/> controls:
-                     <br/> pos. 0,05U GST
+                     <br/> pos. 0,05 U GST
                      <br/> empty
                      <br/> untrans. WT
                      <br/>
-                     <br/> 3,4ml assaycocktail: 170µl CDNB + 170µl GSH + 3060µl PBS
+                     <br/> 3,4 ml master mix: 170 µl CDNB + 170 µl GSH + 3060 µl PBS
                      <br/>
                      <br/>
@@ Line 1,979: / Line 2,034: @@
                      <br/> </p>
              </div>
-        </div>
+             <h3 class="sectionedit90"><a name="shampoo2" id="shampoo2">Shampoo</a></h3>
-        <!-- para20 -->
+             <div class="level3_1">
-    </div>
+                 <p> tested: aGFPnano-CotG(151G) spores </p>
-    <!-- color -->
-    <div class="color12">
-        <div class="para_20">
-            <!-- EDIT85 SECTION "GST-assay" [42228-42583] -->
-             <h3 class="sectionedit86"><a name="section101016" id="section101016">10.10.16</a></h3>
-             <div class="level2"> </div>
-            <!-- EDIT86 SECTION "10.10.16" [42584-42604] -->
-            <h4 class="sectionedit87"><a name="gst-assay5" id="gst-assay5">GST-assay</a></h4>
-            <div class="level3">
-                 <p> measured <em>E. coli</em> lysates again with only one concentration (600µg)
-                    <br/> controls:
-                    <br/> pos. 0,05U GST
-                    <br/> empty
-                    <br/> untrans. WT
-                    <br/>
-                    <br/>
-                    <br/> </p>
              </div>
-             <h4><a name="results12" id="results12">Results</a></h4>
+             <h4><a name="plate_preparation1" id="plate_preparation1">Plate preparation</a></h4>
              <div class="level4">
-                 <p> see above 06.10.16
+                 <p> coated 68 wells with GFP, coating concentration GFP 20 µg/ml and coating volume 100 µl
-                     <br/>
+                    <br/> incubated 2 h at room temerature
+                    <br/> washed twice with 200 µl PBS
+                    <br/> blocked with 200 µl 5% nonfat dry milk in PBS
+                    <br/> incubated for 2 h at roomtemperatur
+                    <br/> washed twice with PBS
+                     <br/> measured the fluorescence of coated GFP
                      <br/>
                      <br/> </p>
              </div>
-             <!-- EDIT87 SECTION "GST-assay" [42605-42798] -->
+             <h4><a name="washing2" id="washing2">Washing</a></h4>
-             <h3 class="sectionedit88"><a name="section111016" id="section111016">11.10.16</a></h3>
+             <div class="level4">
-            <div class="level2"> </div>
+                <p> used 200 µl washing of solutions:
-            <!-- EDIT88 SECTION "11.10.16" [42799-42819] -->
+                    <br/> </p>
-            <h4 class="sectionedit89"><a name="gst-assay6" id="gst-assay6">GST-assay</a></h4>
+                <ul>
-            <div class="level3">
+                    <li class="level1">
-                <p> measured GST-CgeA(152)- &amp; CotG-GST(156)-construct spores
+                        <div class="li"> SDS: 1%, 0.75%, 0.5%, 0.25% (with spores, nanobody and only SDS)</div>
-                     <br/> conditions like on 09.10.16
+                    </li>
-                     <br/>
+                    <li class="level1">
+                        <div class="li"> Tween20: 22%, 24%, 26%, 28% (with spores, nanobody and only Tween20)</div>
+                    </li>
+                    <li class="level1">
+                        <div class="li"> SH1: 12%, 14%, 16%, 18% (with spores, nanobody and only SH1)</div>
+                    </li>
+                    <li class="level1">
+                        <div class="li"> Controls: nanobody in PBS, WT - spore in PBS, construct spore (151G) in PBS, 2% SDS &amp; PBS</div>
+                    </li>
+                </ul>
+                <p>
+                     <br/> incubated wash solutions on the plate for 5 min
+                     <br/> shook for 5 min at 300 rpm
+                    <br/> washed twice with PBS
+                    <br/> measured the emission of remaining GFP
                      <br/>
-                    <br/> <a href="https://static.igem.org/mediawiki/2016/e/e2/T--Freiburg--LabJournal_g5_xlsx46.xlsx" class="media mediafile mf_xlsx" title="labor:11.10.16_gst-assay_spores_settings.xlsx">11.10.16_gst-assay_spores_settings.xlsx</a>
                      <br/>
                      <br/> </p>
              </div>
-             <h4><a name="results13" id="results13">Results</a></h4>
+             <h4><a name="results14" id="results14">Results</a></h4>
              <div class="level4">
-                 <p> <a href="https://static.igem.org/mediawiki/2016/8/81/T--Freiburg--LabJournal_g5_xlsx47.xlsx" class="media mediafile mf_xlsx" title="labor:11.10.16_gst-assay_spores_master.xlsx">11.10.16_gst-assay_spores_master.xlsx</a>
+                 <p> <a href="https://static.igem.org/mediawiki/2016/f/f3/T--Freiburg--deliveryjournal5.xlsx" class="media mediafile mf_xlsx" title="labor:bba_k2114009_agfpnano_ha_ahelix_cotg_in_delta_cotg.xlsx">bba_k2114009_agfpnano_ha_ahelix_cotg_in_delta_cotg.xlsx</a>
-                    <br/>
-                    <br/> The CotG-GST spores show a difference in the slope compared to the WT spores. This difference is significant.
                      <br/>
                      <br/>
                      <br/> </p>
-                <div class="tags"><span>
-</span></div>
              </div>
+            <!-- para20 -->
          </div>
-         <!-- para20 -->
+         <!-- color -->
-    </div>
+        <div class="color12">
-    <!-- color -->
+            <div class="para_20">
+                <!-- EDIT85 SECTION "GST-assay" [42228-42583] -->
+                <h3 class="sectionedit86"><a name="section101016" id="section101016">10.10.16</a></h3>
+                <div class="level2"> </div>
+                <!-- EDIT86 SECTION "10.10.16" [42584-42604] -->
+                <h4 class="sectionedit87"><a name="gst-assay5" id="gst-assay5">GST-assay</a></h4>
+                <div class="level3_1">
+                    <p> measured <em>E. coli</em> lysates again with only one concentration (600 µg)
+                        <br/> controls:
+                        <br/> pos. 0,05 U GST
+                        <br/> empty
+                        <br/> untrans. WT
+                        <br/>
+                        <br/>
+                        <br/> </p>
+                </div>
+                <h4><a name="results12" id="results12">Results</a></h4>
+                <div class="level4">
+                    <p> <a href="https://static.igem.org/mediawiki/2016/5/5a/T--Freiburg--LabJournal_g5_xlsx44.xlsx" class="media mediafile mf_xlsx" title="labor:06.10.16_10.10.16_gst-assay_lysates_master.xlsx">06.10.16_10.10.16_gst-assay_lysates_master.xlsx</a>
+                        <center><img class="something" src="https://static.igem.org/mediawiki/2016/e/e9/T--Freiburg--deliveryjournal1.png"> </center>
+                        <br/>
+                        <br/>
+                        <br/> </p>
+                </div>
+                <h3 class="sectionedit93"><a name="shampoo3" id="shampoo3">Shampoo</a></h3>
+                <div class="level3_1">
+                    <p> tested: aGFPnano-CotZ(157Z) spores </p>
+                </div>
+                <h4><a name="plate_preparation2" id="plate_preparation2">Plate preparation</a></h4>
+                <div class="level4">
+                    <p> coated 68 wells with GFP, coating concentration GFP 20 µg/ml and coating volume 100 µl
+                        <br/> incubated 2 h at room temerature
+                        <br/> washed twice with 200 µl PBS
+                        <br/> blocked with 200 µl 5% nonfat dry milk in PBS
+                        <br/> incubated for 2 h at roomtemperatur
+                        <br/> washed twice with PBS
+                        <br/> measured the fluorescence of coated GFP
+                        <br/>
+                        <br/> </p>
+                </div>
+                <h4><a name="washing3" id="washing3">Washing</a></h4>
+                <div class="level4">
+                    <p> used 200 µl washing of solutions:
+                        <br/> </p>
+                    <ul>
+                        <li class="level1">
+                            <div class="li"> SDS: 1%, 0.75%, 0.5%, 0.25% (with spores, nanobody and only SDS)</div>
+                        </li>
+                        <li class="level1">
+                            <div class="li"> Tween20: 22%, 24%, 26%, 28% (with spores, nanobody and only Tween20)</div>
+                        </li>
+                        <li class="level1">
+                            <div class="li"> SH1: 12%, 14%, 16%, 18% (with spores, nanobody and only SH1)</div>
+                        </li>
+                        <li class="level1">
+                            <div class="li"> Controls: nanobody in PBS, WT - spore in PBS, construct spore (157Z) in PBS, 2% SDS &amp; PBS</div>
+                        </li>
+                    </ul>
+                    <p>
+                        <br/> incubated wash solutions on the plate for 5 min
+                        <br/> shook for 5 min at 300 rpm
+                        <br/> washed twice with PBS
+                        <br/> measured the emission of remaining GFP
+                        <br/>
+                        <br/>
+                        <br/> </p>
+                </div>
+                <h4><a name="results16" id="results16">Results</a></h4>
+                <div class="level4">
+                    <p> <a href="https://static.igem.org/mediawiki/2016/d/df/T--Freiburg--deliveryjournal6.xlsx" title="labor:bba_k2114001_agfpnano_ha_ahelix_cotz_in_δcotz_spore.xlsx">bba_k2114001_agfpnano_ha_ahelix_cotz_in_δcotz_spore.xlsx</a>
+                        <br/>
+                        <br/>
+                        <br/> </p>
+                    <h3 class="sectionedit88"><a name="section111016" id="section111016">11.10.16</a></h3>
+                    <div class="level2"> </div>
+                    <h3 class="sectionedit95"><a name="shampoo4" id="shampoo4">Shampoo</a></h3>
+                    <div class="level3_1">
+                        <p> prepared 3 plates
+                            <br/> tested: aGFPnano-CotZ(159WT), aGFPnano-CotZ(157WT), aGFPnano-CotG(151WT) </p>
+                    </div>
+                    <h4><a name="plate_preparation3" id="plate_preparation3">Plate preparation</a></h4>
+                    <div class="level4">
+                        <p> coated 68 wells with GFP, coating concentration GFP 20 µg/ml and coating volume 100 µl
+                            <br/> incubated 2 h at room temerature
+                            <br/> washed twice with 200 µl PBS
+                            <br/> blocked with 200 µl 5% nonfat dry milk in PBS
+                            <br/> incubated for 2 h at roomtemperatur
+                            <br/> washed twice with PBS
+                            <br/> measured the fluorescence of coated GFP
+                            <br/>
+                            <br/> </p>
+                    </div>
+                    <h4><a name="washing4" id="washing4">Washing</a></h4>
+                    <div class="level4">
+                        <p> used 200 µl washing of solutions:
+                            <br/> </p>
+                        <ul>
+                            <li class="level1">
+                                <div class="li"> SDS: 1%, 0.75%, 0.5%, 0.25% (with spores, nanobody and only SDS)</div>
+                            </li>
+                            <li class="level1">
+                                <div class="li"> Tween20: 22%, 24%, 26%, 28% (with spores, nanobody and only Tween20)</div>
+                            </li>
+                            <li class="level1">
+                                <div class="li"> SH1: 12%, 14%, 16%, 18% (with spores, nanobody and only SH1)</div>
+                            </li>
+                            <li class="level1">
+                                <div class="li"> Controls: nanobody in PBS, WT - spore in PBS, construct spore in PBS, 2% SDS &amp; PBS</div>
+                            </li>
+                        </ul>
+                        <p>
+                            <br/> incubated wash solutions on the plate for 5 min
+                            <br/> shook for 5 min at 300 rpm
+                            <br/> washed twice with PBS
+                            <br/> measured the emission of remaining GFP
+                            <br/>
+                            <br/>
+                            <br/> </p>
+                    </div>
+                    <h4><a name="results17" id="results17">Results</a></h4>
+                    <div class="level4">
+                        <p> <a href="https://static.igem.org/mediawiki/2016/0/04/T--Freiburg--deliveryjournal7.xlsx" title="labor:bba_k2114001_agfpnano_ha_ahelix_cotz_in_wt_spores.xlsx">bba_k2114001_agfpnano_ha_ahelix_cotz_in_wt_spores.xlsx</a>
+                            <br/> <a href="https://static.igem.org/mediawiki/2016/9/9b/T--Freiburg--deliveryjournal8.xlsx" title="labor:bba_k2114002_agfpnano_ha_g4s_cotz_in_wt_.xlsx">bba_k2114002_agfpnano_ha_g4s_cotz_in_wt_.xlsx</a>
+                            <br/> <a href="https://static.igem.org/mediawiki/2016/e/e7/T--Freiburg--deliveryjournal9.xlsx" title="labor:bba_k2114009_agfpnano_ha_ahelix_cotg_in_wt.xlsx">bba_k2114009_agfpnano_ha_ahelix_cotg_in_wt.xlsx</a>
+                            <br/>
+                            <br/>
+                            <br/> </p>
+                        <h4 class="sectionedit89"><a name="gst-assay6" id="gst-assay6">GST-assay</a></h4>
+                        <div class="level3_1">
+                            <p> measured GST-CgeA(152)- &amp; CotG-GST(156)-construct spores
+                                <br/> conditions like on 09.10.16
+                                <br/>
+                                <br/>
+                                <br/> <a href="https://static.igem.org/mediawiki/2016/e/e2/T--Freiburg--LabJournal_g5_xlsx46.xlsx" class="media mediafile mf_xlsx" title="labor:11.10.16_gst-assay_spores_settings.xlsx">11.10.16_gst-assay_spores_settings.xlsx</a>
+                                <br/>
+                                <br/> </p>
+                        </div>
+                        <h4><a name="results13" id="results13">Results</a></h4>
+                        <div class="level4">
+                            <p> <a href="https://static.igem.org/mediawiki/2016/8/81/T--Freiburg--LabJournal_g5_xlsx47.xlsx" class="media mediafile mf_xlsx" title="labor:11.10.16_gst-assay_spores_master.xlsx">11.10.16_gst-assay_spores_master.xlsx</a>
+                                <br/>
+                                <center><img class="something" src="https://static.igem.org/mediawiki/2016/7/7e/T--Freiburg--deliveryjournal2.png"> </center>
+                                <br/> The CotG-GST spores(156) show a difference in the slope compared to the WT spores. This difference is significant.
+                                <br/>
+                                <br/>
+                                <br/>
+                                <center><img class="something" src="https://static.igem.org/mediawiki/2016/9/9c/T--Freiburg--deliveryjournal3.png"> </center>
+                                <br/>
+                                <br/> </p>
+                            <div class="tags"><span>
+</span></div>
+                        </div>
+                    </div>
+                    <!-- para20 -->
+                </div>
+                <!-- color -->
 </body>
 </html>
 {{Freiburg/Footer}}