Difference between revisions of "Team:SDU-Denmark/Notebook"

Revision as of 13:05, 18 October 2016

Notebook

Week 10

Week 13

Week 14

Week 15

We learned how we find thing in the lab, and tried to make some basic experiments. On the photo is Joel learning how to pipet

Week 17

This year the faculty of Biochemistry and Molecular Biology had a big workshop area with the theme “The Good, the Bad and the Ugly”: the probiotic bacteria, the pathogenic bacteria and the multiresistant bacteria.

Week 18

Week 24

Week 26

The brick was ordered from IDT and received: The received gblock had a weight of 200 ng. and was resuspended in dH2O. 5 µl diluted gblock and 95 µl ddH2O to make a 20X dilution.

What we have done with the BioBrick K2018043 can be read in the protocol: Cloning Basic part K2018043 into pSB1C3

The brick was ordered from IDT and received: The received gblock had a weight of 200 ng. and was resuspended in dH2O. 5 µl diluted gblock and 95 µl ddH2O to make a 20X dilution.

What we have done with the BioBrick K2018044 can be read in the protocol: Cloning Basic part K2018044 into pSB1C3

The brick was ordered from IDT and received: The received gblock had a weight of 200 ng. and was resuspended in dH2O. 5 µl diluted gblock and 95 µl ddH2O to make a 20X dilution.

What we had done with the BioBrick K1763002 can be read in the protocol: Cloning Basic part K1763002 into pSB1C3

The brick was ordered from IDT and received: The received gblock had a weight of 200 ng. and was resuspended in dH2O. 5 µl diluted gblock and 95 µl ddH2O to make a 20X dilution.

What we had done with the BioBrick K1763003 can be read in the protocol: Cloning Basic part K1763003 into pSB1C3

The brick was ordered from IDT and received: The received gblock had a weight of 200 ng. and was resuspended in dH2O. 5 µl diluted gblock and 95 µl ddH2O to make a 20X dilution.

What we had done with the BioBrick K1763004 can be read in the protocol: Cloning Basic part K1763004 into pSB1C3

The brick was ordered from IDT and received: The received gblock had a weight of 200 ng. and was resuspended in dH2O. 5 µl diluted gblock and 95 µl ddH2O to make a 20X dilution.

What we had done with the BioBrick K1763009 can be read in the protocol: Cloning Basic part K1763009 into pSB1C3

The brick was ordered from IDT and received: The received gblock had a weight of 200 ng. and was resuspended in dH2O. 5 µl diluted gblock and 95 µl ddH2O to make a 20X dilution.

What we had done with the BioBrick K1763010 can be read in the protocol: Cloning Basic part K1763010 into pSB1C3

The brick was ordered from IDT and received: The received gblock had a weight of 200 ng. and was resuspended in dH2O. 5 µl diluted gblock and 95 µl ddH2O to make a 20X dilution.

What we had done with the BioBrick K1763011 can be read in the protocol: Cloning Basic part K1763011 into pSB1C3

The brick was ordered from IDT and received: The received gblock had a weight of 200 ng. and was resuspended in dH2O. 5 µl diluted gblock and 95 µl ddH2O to make a 20X dilution.

What we had done with the BioBrick K1763012 can be read in the protocol: Cloning Basic part K1763012 into pSB1C3

Week 27

The BioBrick was ordered from IDT in a plasmid pUCIDT-AMP and after receiving it, it was resuspended to a final concentration of 0.1 µg/µl.

After the resuspension, the plasmid was transformed into Top10 E. coli by following SOP0023_v01 Ca2+ Transformation , with the following remarks:

2.5 µl plasmid was used instead of the normal 1 µl.

What we had done with the BioBrick K2018011 can be read in the protocols: Cloning composite part K2018011 into pSB1C3, Cloning ThuricinS with DA silk-overhangs into pSB1C3, Cloning ThuricinS with DE silk-overhangs into pSB1C3, Cloning ThuricinS with EA silk-overhangs into pSB1C3, Cloning ThuricinS with ED silk-overhangs into pSB1C3 and IMPACT Purification of K2018011

Week 28

The BioBrick was ordered from IDT in a plasmid pUCIDT-AMP and after receiving it, it was resuspended to a final concentration of 0.1 µg/µl.

After the resuspension, the plasmid was transformed into Top10 E. coli by following SOP0023_v01 Ca2+ Transformation Ca2+ Transformation , with the following remarks:

2.5 µl plasmid was used instead of the normal 1 µl.

What we had done with the BioBrickK2018010 can be read in the protocols: Cloning composite part K2018010 into pSB1C3. and IMPACT Purification of K2018010

The BioBrick containing the gene K2018011 was digested with PstI and XbaI overnight by using SOP0017 .

After digesting, the sample was ligated by following SOP0015 with the standard iGEM plasmid, pSB1C3, overnight, and transformed into E.coli Top10.

cPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	98 °C	2 min
2	34 cycles	98 °C	10 sec
		72 °C	15 sec
		72 °C	>15 sec
3	Final extension	72 °C	5 min
4	Keep the sample cold	12 °C	HOLD

After a colony PCR, SOP0021 , was run and verified on a gel, the following result was given: Our part is 259 bp and primer overhangs are 150 X 2 bp, which is, in total, 559 bp: A gel electrophoresis from cPCR showed a band around 600 bp, so we assumed that we had the correct part inserted into the iGEM standard plasmid.

What we had done with the BioBrick K2018011 can be read in the protocols: Cloning composite part K2018011 into pSB1C3, Cloning ThuricinS with DA silk-overhangs into pSB1C3, Cloning ThuricinS with DE silk-overhangs into pSB1C3, Cloning ThuricinS with EA silk-overhangs into pSB1C3, Cloning ThuricinS with ED silk-overhangs into pSB1C3 and IMPACT Purification of K2018011

The BioBrick was ordered from IDT in a plasmid pUCIDT-AMP and after receiving it, it was resuspended to a final concentration of 0.1 µg/µl.

After the resuspension, the plasmid was transformed into Top10 E. coli by following SOP0023_v01 Ca2+ Transformation , with the following remarks:

2.5 µl plasmid was used instead of the normal 1 µl.

What we had done with the BioBrickK2018014 can be read in the protocols: Cloning composite part K2018014 into pSB1C3 and IMPACT Purification of K2018014

The BioBrick was ordered from IDT in a plasmid pUCIDT-AMP and after receiving it, it was resuspended to a final concentration of 0.1 µg/µl.

After the resuspension, the plasmid was transformed into Top10 E. coli by following SOP0023_v01 Ca2+ Transformation , with the following remarks:

2.5 µl plasmid was used instead of the normal 1 µl.

What we had done with the BioBrickK2018019 can be read in the protocol: Cloning composite part K2018019 into pSB1C3

Week 29

The BioBrick was first amplified by phusion PCR, SOP0010 , thereafter digested with restrictions enzymes, following SOP0017 . The restriction enzymes were: PstI for 2 h and EcoRI for 30 min. After digesting, the sample was verified by running a gel electrophoresis.

pPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	95 °C	3 min
2	34 cycles	95 °C	25 sec
		58 °C	25 sec
		72 °C	30 sec
3	Final extension	72 °C	1 min
4	Keep the sample cold	20 °C	HOLD

In the next step, the vector was ligated with the insert, SG47, overnight by following SOP0015 , and thereafter it was transformed, following SOP0009, into E.coli Top10. The ratio of the ligation is shown hereunder:

Ratio	1:2	1:5
Ligase Buffer	2 µl	2 µl
Ligase	1 µl	1 µl
SG47	2 µl	5 µl
SG58	1 µl	1 µl
ddH2O	14 µl	11 µl

To verify that the transformation and ligation succeeded, a colony PCR was run, SOP0021 .

cPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	95 °C	3 min
2	34 cycles	95 °C	25 sec
		58 °C	25 sec
		72 °C	30 sec
3	Final extension	72 °C	1 min
4	Keep the sample cold	20 °C	HOLD

The Gel Photo below shows the cPCR results. VR and VF2 primers adds a total of 300 bp, the gene is 119 bp which gives a total of DNA length of 419 bp. From this gel, the ligation of the gen incorporated into psB1C3 can be observed .

Picture of cPCR of different colonies on the same plate (each gene had its own plate). All the colony PCR shows a band near the 4-500 bp band, which was desired.

The second gel photo shows cutting of SB71 with Pst1 at 2 hours with 37 °C and EcoR1 at 30 minutes. Every second well had added restriction enzymes.If a band of approximately 160 bp could be seen on the gel, it was concluded as being successful and sent to sequencing.

What we had done with the BioBrick K1763002 can be read in the protocol: Cloning Basic part K1763002 into pSB1C3

The BioBrick was first amplified by phusion PCR, SOP0010 , and thereafter digested with restrictions enzymes, following SOP0017 . The restriction enzymes were: PstI for 2 h and EcoRI for 30 min. After digesting, the sample was verified by running a gel electrophoresis.

pPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	95 °C	3 min
2	34 cycles	95 °C	25 sec
		58 °C	25 sec
		72 °C	30 sec
3	Final extension	72 °C	1 min
4	Keep the sample cold	20 °C	HOLD

In the next step, the vector was ligated with the insert, SG48, overnight by following SOP0015 , and thereafter it was transformed, following SOP0009, into E.coli Top10. The ratio of the ligation is shown hereunder:

Ratio	1:2	1:5
Ligase Buffer	2 µl	2 µl
Ligase	1 µl	1 µl
SG48	2 µl	5 µl
SG58	1 µl	1 µl
ddH2O	14 µl	11 µl

To verify that the transformation and ligation succeeded, a colony PCR was run, SOP0021 .

cPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	95 °C	3 min
2	34 cycles	95 °C	25 sec
		58 °C	25 sec
		72 °C	30 sec
3	Final extension	72 °C	1 min
4	Keep the sample cold	20 °C	HOLD

The Gel Photo below shows the cPCR results. VR and VF2 primers adds a total of 300 bp, the gene is 119 bp which gives a total of DNA length of 419 bp. From this gel, the ligation of the gen incorporated into psB1C3 can be observed .

Picture of cPCR of different colonies on the same plate (each gene had its own plate). All the colony PCR shows a band near the 4-500 bp band.

The second gel photo shows cutting of SB73 with Pst1 at 2 hours with 37 °C and EcoR1 at 30 minutes. Every second well had added restriction enzymes.If a band of approximately 160 bp could be seen on the gel, it was concluded as being successful and sent to sequencing.

What we had done with the BioBrick K1763003 can be read in the protocol: Cloning Basic part K1763003 into pSB1C3

The BioBrick was first amplified by phusion PCR, SOP0010 , and thereafter digested with restrictions enzymes, following SOP0017 . The restriction enzymes were: PstI for 2 h and EcoRI for 30 min. After digesting, the sample was verified by running a gel electrophoresis.

pPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	95 °C	3 min
2	34 cycles	95 °C	25 sec
		58 °C	25 sec
		72 °C	30 sec
3	Final extension	72 °C	1 min
4	Keep the sample cold	20 °C	HOLD

In the next step, the vector was ligated with the insert, SG49, overnight by following SOP0015 , and thereafter it was transformed, following SOP0009, into E.coli Top10. The ratio of the ligation is shown hereunder:

Ratio	1:2	1:5
Ligase Buffer	2 µl	2 µl
Ligase	1 µl	1 µl
SG49	2 µl	5 µl
SG58	1 µl	1 µl
ddH2O	14 µl	11 µl

To verify that the transformation and ligation succeeded, a colony PCR was run, SOP0021 .

cPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	95 °C	3 min
2	34 cycles	95 °C	25 sec
		58 °C	25 sec
		72 °C	30 sec
3	Final extension	72 °C	1 min
4	Keep the sample cold	20 °C	HOLD

The Gel Photo below shows the cPCR results. VR and VF2 primers adds a total of 300 bp, the gene is 119 bp which gives a total of DNA length of 419 bp. From this gel, the ligation of the gen incorporated into psB1C3 can be seen.

Picture of cPCR of different colonies on the same plate (each gene had its own plate). All the colony PCR shows a band near the 4-500 bp band.

The second gel photo shows cutting of SB75 with Pst1 at 2 hours with 37 °C and EcoR1 at 30 minutes. Every second well had added restriction enzymes.If a band of approximately 160 bp could be seen on the gel, it was concluded as being successful and sent to sequencing.

What we had done with the BioBrick K1763004 can be read in the protocol: Cloning Basic part K1763004 into pSB1C3

The BioBrick was first amplified by phusion PCR, SOP0010 , and thereafter digested with restrictions enzymes, following SOP0017 . The restriction enzymes were: PstI for 2 h and EcoRI for 30 min. After digesting, the sample was verified by running a gel electrophoresis.

pPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	95 °C	3 min
2	34 cycles	95 °C	25 sec
		58 °C	25 sec
		72 °C	>30 sec
3	Final extension	72 °C	1 min
4	Keep the sample cold	20 °C	HOLD

In the next step, the vector was ligated with the insert, SG56, overnight by following SOP0015 , and thereafter it was transformed, following SOP0009, into E.coli Top10. The ratio of the ligation is shown hereunder:

Ratio	1:2	1:5
Ligase Buffer	2 µl	2 µl
Ligase	1 µl	1 µl
SG56	2 µl	5 µl
SG58	1 µl	1 µl
ddH2O	14 µl	11 µl

To verify that the transformation and ligation succeeded, a colony PCR was run, SOP0021 .

cPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	95 °C	3 min
2	34 cycles	95 °C	25 sec
		58 °C	25 sec
		72 °C	30 sec
3	Final extension	72 °C	1 min
4	Keep the sample cold	20 °C	HOLD

The Gel Photo below shows the cPCR results. VR and VF2 primers adds a total of 300 bp, the gene is 119 bp which gives a total of DNA length of 419 bp. From this gel, the ligation of the gen incorporated into psB1C3 can be observed .

Picture of cPCR of different colonies on the same plate (each gene had its own plate). All the colony PCR shows a band near the 4-500 bp band.

The second gel photo shows cutting of SB77 with Pst1 at 2 hours with 37 °C and EcoR1 at 30 minutes. Every second well had added restriction enzymes.If a band of approximately 160 bp could be seen on the gel, it was concluded as being successful and sent to sequencing.

What we had done with the BioBrick K1763009 can be read in the protocol: Cloning Basic part K1763009 into pSB1C3

The BioBrick was first amplified by phusion PCR, SOP0010 , and thereafter digested with restrictions enzymes, following SOP0017 . The restriction enzymes were: PstI for 2 h and EcoRI for 30 min. After digesting, the sample was verified by running a gel electrophoresis.

pPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	95 °C	3 min
2	34 cycles	95 °C	25 sec
		58 °C	25 sec
		72 °C	>30 sec
3	Final extension	72 °C	1 min
4	Keep the sample cold	20 °C	HOLD

In the next step, the vector was ligated with the insert, SG43, overnight by following SOP0015 , and thereafter it was transformed, following SOP0009, into E.coli Top10. The ratio of the ligation is shown hereunder:

Ratio	1:2	1:5
Ligase Buffer	2 µl	2 µl
Ligase	1 µl	1 µl
SG43	2 µl	5 µl
SG58	1 µl	1 µl
ddH2O	14 µl	11 µl

To verify that the transformation and ligation succeeded, a colony PCR was run, SOP0021 .

cPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	95 °C	3 min
2	34 cycles	95 °C	25 sec
		58 °C	25 sec
		72 °C	30 sec
3	Final extension	72 °C	1 min
4	Keep the sample cold	20 °C	HOLD

The Gel Photo below shows the cPCR results. VR and VF2 primers adds a total of 300 bp, the gene is 119 bp which gives a total of DNA length of 419 bp. From this gel, the ligation of the gen incorporated into psB1C3 can be observed.

Picture of cPCR of different colonies on the same plate (each gene had its own plate). All the colony PCR shows a band near the 4-500 bp band.

The second gel photo shows cutting of SB63 with Pst1 at 2 hours with 37 °C and EcoR1 at 30 minutes. Every second well had added restriction enzymes.If a band of approximately 160 bp could be seen on the gel, it was concluded as being successful and sent to sequencing.

What we had done with the BioBrick K1763010 can be read in the protocol: Cloning Basic part K1763010 into pSB1C3

The BioBrick was first amplified by phusion PCR, SOP0010 , and thereafter digested with restrictions enzymes, following SOP0017 . The restriction enzymes were: PstI for 2 h and EcoRI for 30 min. After digesting, the sample was verified by running a gel electrophoresis.

pPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	95 °C	3 min
2	34 cycles	95 °C	25 sec
		58 °C	25 sec
		72 °C	>30 sec
3	Final extension	72 °C	1 min
4	Keep the sample cold	20 °C	HOLD

In the next step, the vector was ligated with the insert, SG44, overnight by following SOP0015 , and thereafter it was transformed, following SOP0009, into E.coli Top10. The ratio of the ligation is shown hereunder:

Ratio	1:2	1:5
Ligase Buffer	2 µl	2 µl
Ligase	1 µl	1 µl
SG44	2 µl	5 µl
SG58	1 µl	1 µl
ddH2O	14 µl	11 µl

To verify that the transformation and ligation succeeded, a colony PCR was run, SOP0021 .

cPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	95 °C	3 min
2	34 cycles	95 °C	25 sec
		58 °C	25 sec
		72 °C	30 sec
3	Final extension	72 °C	1 min
4	Keep the sample cold	20 °C	HOLD

The Gel Photo below shows the cPCR results. VR and VF2 primers adds a total of 300 bp, the gene is 119 bp which gives a total of DNA length of 419 bp. From this gel, the ligation of the gen incorporated into psB1C3 can be observed .

Picture of cPCR of different colonies on the same plate (each gene had its own plate). All the colony PCR shows a band near the 4-500 bp band.

The second gel photo shows cutting of SB65 with Pst1 at 2 hours with 37 °C and EcoR1 at 30 minutes. Every second well had added restriction enzymes.If a band of approximately 160 bp could be seen on the gel, it was concluded as being successful and sent to sequencing.

What we had done with the BioBrick K1763011 can be read in the protocol: Cloning Basic part K1763011 into pSB1C3

The BioBrick was first amplified by phusion PCR, SOP0010 , and thereafter digested with restrictions enzymes, following SOP0017 . The restriction enzymes were: PstI for 2 h and EcoRI for 30 min. After digesting, the sample was verified by running a gel electrophoresis.

pPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	95 °C	3 min
2	34 cycles	95 °C	25 sec
		58 °C	25 sec
		72 °C	>30 sec
3	Final extension	72 °C	1 min
4	Keep the sample cold	20 °C	HOLD

In the next step, the vector was ligated with the insert, SG45, overnight by following SOP0015 , and thereafter it was transformed, following SOP0009, into E.coli Top10. The ratio of the ligation is shown hereunder:

Ratio	1:2	1:5
Ligase Buffer	2 µl	2 µl
Ligase	1 µl	1 µl
SG45	2 µl	5 µl
SG58	1 µl	1 µl
ddH2O	14 µl	11 µl

To verify that the transformation and ligation succeeded, a colony PCR was run, SOP0021 .

cPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	95 °C	3 min
2	34 cycles	95 °C	25 sec
		58 °C	25 sec
		72 °C	30 sec
3	Final extension	72 °C	1 min
4	Keep the sample cold	20 °C	HOLD

The Gel Photo below shows the cPCR results. VR and VF2 primers adds a total of 300 bp, the gene is 119 bp which gives a total of DNA length of 419 bp. From this gel, the ligation of the gen incorporated into psB1C3 can be observed .

Picture of cPCR of different colonies on the same plate (each gene had its own plate). All the colony PCR shows a band near the 4-500 bp band.

The second gel photo shows cutting of SB67 with Pst1 at 2 hours with 37 °C and EcoR1 at 30 minutes. Every second well had added restriction enzymes.If a band of approximately 160 bp could be seen on the gel, it was concluded as being successful and sent to sequencing.

What we had done with the BioBrick K1763012 can be read in the protocol: Cloning Basic part K1763012 into pSB1C3

Week 30

The BioBrick containing the gene K2018010 was digested with PstI overnight and with EcoRI the day after by using SOP0017 .

After digesting, the sample was verified by gel electrophoresis. The result gave the following: The Gel Photo below shows BG51 (At the photo named; 51). The composite part is 395 bp long. It is possible from the Gel photo to assume that the DNA is cut correctly. From this gel, is the BG51 band of approximately 400 bp purified and thereafter ligated into the standard iGEM plasmid.

After digesting, the sample was ligated by following SOP0015 , with the standard iGEM plasmid, pSB1C3, overnight, and transformed into E.coli Top10.

cPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	98 °C	2 min
2	34 cycles	98 °C	10 sec
		72 °C	15 sec
		72 °C	15 sec
3	Final extension	72 °C	5 min
4	Keep the sample cold	12 °C	HOLD

Hereafter a colony PCR was runned and verified on a gel, the result is given below:

The Gel Photo below shows to the left the ladder (green). The following bands are BB54, 1, 2 and 4 which are the ligation of the vector PR17 and the insert BG51. From the photo, it can be seen that sample 54.1 and 54.3 are the vector with the insert, and 54.2 is a religation.

What we had done with the BioBrickK2018010 can be read in the protocols: Cloning composite part K2018010 into pSB1C3. and IMPACT Purification of K2018010

The BioBrick containing the gene K2018014 was digested with PstI overnight and with EcoRI the day after by using SOP0017 .

After digesting, the sample was verified by gel electrophoresis. The result gave the following: The Gel Photo below shows BG52 (At the photo named; 52).The composite part is 458 bp long. It is possible, from the Gel photo, to assume that the DNA is cut correctly. From this gel, the BG52 band of approximately 400 bp purified and thereafter ligated into the standard iGEM plasmid.

After digesting, the sample was ligated by following SOP0015 , with the standard iGEM plasmid, pSB1C3, overnight, and transformed into E.coli Top10.

Ratio	1:2	1:5	1:10
Ligase Buffer	2 µl	2 µl	2 µl
Ligase	1 µl	1 µl	1µl
BG75	2 µl	5 µl	10 µl
BG17	1 µl	1 µl	1 µl
ddH2O	14 µl	11 µl	6 µl

cPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	98 °C	2 min
2	34 cycles	98 °C	10 sec
		72 °C	15 sec
		72 °C	15 sec
3	Final extension	72 °C	5 min
4	Keep the sample cold	12 °C	HOLD

Hereafter a colony PCR was runned and verified on a gel, the result is given below:

The Gel Photo below shows to the left, the ladder (green). The following bands are BB55, 1, 2 and 4 are the ligation of the vector PR17 and the insert BG52. From the photo, it can be assumed that all 3 samples in BB55 are the vector with insert.

What we had done with the BioBrickK2018014 can be read in the protocols: Cloning composite part K2018014 into pSB1C3 and IMPACT Purification of K2018014

The BioBrick was ordered from IDT in a plasmid pUCIDT-AMP. When received, it was resuspended to a final concentration at 0.1 µg/µl.

After the resuspension, the plasmid is transformed into Top10 E. coli by following SOP0023_v01 Ca2+ Transformation with the following remarks:

2.5 µl plasmid were used instead of the normal 1 µl.

What we had done with the BioBrick K2018012 can be read in the protocols: Cloning composite part K2018012 into pSB1C3 and IMPACT Purification of K2018012

The BioBrick was ordered from IDT in a plasmid pUCIDT-AMP. When received, it was resuspended to a final concentration at 0.1 µg/µl.

After the resuspension, the plasmid is transformed into Top10 E. coli by following SOP0023_v01 Ca2+ Transformation with the following remarks:

2.5 µl plasmid were used instead of the normal 1 µl.

What we had done with the BioBrick K2018015 can be read in the protocols: <Cloning composite part K2018015 into pSB1C3and IMPACT Purification of K2018015

The BioBrick, that was ordered from IDT had a defected promoter, and therefore we replaced the promoter region with a functional one. This was done by a phusion PCR, SOP0010 . The primer was; k2018015_IDT.

ddH2O	32.5 µl
5 x HFbuffter	10 µl
dNTPs	1 µl
Primer; K2018015_IDT	2.5 µl
Suffix-primer	2.5 µl
Polymerase	0.5 µl

pPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	95 °C	2 min
2	34 cycles	95 °C	10 sec
		72 °C	16 sec
		72 °C	15 sec
3	Final extension	72 °C	10 min
4	Keep the sample cold	20 °C	HOLD

The Gel photo below shows the result from the pPCR. This was done to replace a defective promoter, with the correct one. This is done by using the Primer; k2018015_IDT. As it can be seen from the photo, is first the ladder (green) and then sample BR74. The gene is 572 bp long. and it is therefore possible from the gel to assume that the pPCR is succeed.

Note: There is loaded 5 µl of each sample

What we had done with the BioBrick K2018015 can be read in the protocols: <Cloning composite part K2018015 into pSB1C3and IMPACT Purification of K2018015

The BioBrick containing the gene K2018019 was digested with PstI for 2 hours and EcoRI for 30 min. by using SOP0017 .

After digesting, the sample was verified by gel electrophoresis. The result gave the following: The two Gel Photos below show the results for BG59 (At the photo named; 59). The composite part is 1739 bp long. It is possible from the Gel photo to assume that the DNA is cut correctly. From this gel, is the BG59 band of around 2000 bp purified and thereafter ligated into the standard iGEM plasmid.

After digesting, the sample was ligated by following SOP0015 , with the standard iGEM plasmid, pSB1C3, overnight, and thereafter transformed, SOP009_v01, into E.coli Top10.

The ratio of the ligation is shown hereunder:

Ratio	1:0	1:5	1:10
Ligase Buffer	2 µl	2 µl	2 µl
Ligase	1 µl	1 µl	1µl
BG59	0 µl	5 µl	10 µl
BG60	1 µl	1 µl	1 µl
ddH2O	16 µl	11 µl	6 µl

cPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	98 °C	2 min
2	34 cycles	98 °C	10 sec
		72 °C	15 sec
		72 °C	15 sec
3	Final extension	72 °C	5 min
4	Keep the sample cold	12 °C	HOLD

Hereafter a colony PCR was runned and verified on a gel. The result is given in the following section: The Gel Photo below shows to the left the ladder (green). The following bands are BB59, 1 to 14, which represent the ligation of the vector BR60 with the insert BG59. From the photo it can be assumed that all the samples are perfectly ligated. One of the colonies can therefore, be used to make ON for the plasmid purification and Freeze stock.

What we had done with the BioBrickK2018019 can be read in the protocol: Cloning composite part K2018019 into pSB1C3

Week 31

The BioBrick containing the gene K2018012 was digested with PstI and XbaI overnight by using SOP0017 .

After digesting, the sample was ligated by following SOP0015 , with the standard iGEM plasmid, pSB1C3, overnight, and thereafter transformed, SOP009_v01, into E.coli Top10.

The ratio of the ligation is shown hereunder:

Ratio	1:0	1:5	1:10
Ligase Buffer	2 µl	2 µl	2 µl
Ligase	1 µl	1 µl	1µl
BG75	0 µl	5 µl	10 µl
BG17	1 µl	1 µl	1 µl
ddH2O	16 µl	11 µl	6 µl

cPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	98 °C	2 min
2	34 cycles	98 °C	10 sec
		72 °C	15 sec
		72 °C	15 sec
3	Final extension	72 °C	5 min
4	Keep the sample cold	12 °C	HOLD

Hereafter a colony PCR, SOP0021 , was runned and verified on a gel, The result is given below:

The following gel picture shows that we successfully cloned our part K2018012 into the standard iGEM plasmid, pSB1C3.

What we had done with the BioBrick K2018012 can be read in the protocols: Cloning composite part K2018012 into pSB1C3 and IMPACT Purification of K2018012

The BioBrick containing the gene K2018015 was digested with PstI and XbaI overnight by using SOP0017 .

After digesting, the sample was ligated by following SOP0015 , with the standard iGEM plasmid, pSB1C3, overnight, and thereafter transformed, SOP009_v01, into E.coli Top10.

The ratio of the ligation is shown hereunder:

Ratio	1:0	1:5	1:10
Ligase Buffer	2 µl	2 µl	2 µl
Ligase	1 µl	1 µl	1µl
BG78	0 µl	5 µl	10 µl
BG17	1 µl	1 µl	1 µl
ddH2O	16 µl	11 µl	6 µl

cPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	98 °C	2 min
2	34 cycles	98 °C	10 sec
		72 °C	15 sec
		72 °C	15 sec
3	Final extension	72 °C	5 min
4	Keep the sample cold	12 °C	HOLD

After a colony PCR, SOP0021 , was runned and verified on a gel, the results were the following:

The Gel Photos below show the first Gel , to the left the ladder (green) can be seen. The following bands are BB79, 1 to 14. On the other gel can be seen first the ladder (green), and then BB79 sample 15 to 23. BB79 is the ligation of the vector BG17 and the insert BG78. From the photo it can be assumed that all the samples, except; 2 and 6, are perfectly ligated. One of the colonies are therefore used to make ON for plasmid purification and Freeze stock.

What we had done with the BioBrick K2018015 can be read in the protocols: <Cloning composite part K2018015 into pSB1C3and IMPACT Purification of K2018015

Week 32

Week 33

K2018011in standard iGEM plasmid (pSB1C3) was used in a phusion PCR, SOP0010 . The primes have an overhang, which makes the gene compatible with the silk overhang, and thereby enabling us to make a hybrid silk fiber.

pPCR program:

Segment	Step	Temperature	Duration
1	Initial denaturation	98 °C	2 min
2	34 cycles	98 °C	30 sec
		70 °C	30 sec
		72 °C	15 sec
3	Final extension	72 °C	5 min
4	Keep the sample cold	20 °C	HOLD

The result is verified by Gel electrophoresis. The result is shown here:

Lane no. 5 is ThuricinS with DA-overhangs and is expected to be 217 bp long. We have used the yellow ladder, where the fourth band represents 200 bp. Based on that, it can be assumed that we succeeded at getting silk-overhangs on ThuricinS.

What we had done with the BioBrick K2018011 can be read in the protocols: Cloning composite part K2018011 into pSB1C3, Cloning ThuricinS with DA silk-overhangs into pSB1C3, Cloning ThuricinS with DE silk-overhangs into pSB1C3, Cloning ThuricinS with EA silk-overhangs into pSB1C3, Cloning ThuricinS with ED silk-overhangs into pSB1C3 and IMPACT Purification of K2018011

K2018011in standard iGEM plasmid (pSB1C3) was used in a phusion PCR, SOP0010 . The primes have an overhang, which makes the gene compatible with the silk overhang, and thereby enabling us to make a hybrid silk fiber.

pPCR program:

Segment	Step	Temperature	Duration
1	Initial denaturation	98 °C	2 min
2	34 cycles	98 °C	30 sec
		70 °C	30 sec
		72 °C	15 sec
3	Final extension	72 °C	5 min
4	Keep the sample cold	20 °C	HOLD

The result is verified by Gel electrophoresis. The result is shown here:

The lane marked no. 2 is ThuricinS with DE-overhangs and it is assumed to be 217 bp long. We have used the yellow ladder, where the fourth band is 200 bp long. Based on that, it can be assumed that we succeeded at getting silk-overhangs on ThuricinS.

What we had done with the BioBrick K2018011 can be read in the protocols: Cloning composite part K2018011 into pSB1C3, Cloning ThuricinS with DA silk-overhangs into pSB1C3, Cloning ThuricinS with DE silk-overhangs into pSB1C3, Cloning ThuricinS with EA silk-overhangs into pSB1C3, Cloning ThuricinS with ED silk-overhangs into pSB1C3 and IMPACT Purification of K2018011

K2018011in standard iGEM plasmid (pSB1C3) was used in a phusion PCR, SOP0010 . The primes have an overhang, which makes the gene compatible with the silk overhang, and thereby enabling us to make a hybrid silk fiber.

pPCR program:

Segment	Step	Temperature	Duration
1	Initial denaturation	98 °C	2 min
2	34 cycles	98 °C	30 sec
		70 °C	30 sec
		72 °C	15 sec
3	Final extension	72 °C	5 min
4	Keep the sample cold	20 °C	HOLD

The result is verified by Gel electrophoresis. The result is shown here:

The lane marked no. 7 and 8 are ThuricinS with EA-overhangs and it are expected to be 217 bp long. We have used the yellow ladder, where the fourth band is 200 bp long. Based on that, it can be expected that we succeeded at getting silk-overhangs on ThuricinS.

What we had done with the BioBrick K2018011 can be read in the protocols: Cloning composite part K2018011 into pSB1C3, Cloning ThuricinS with DA silk-overhangs into pSB1C3, Cloning ThuricinS with DE silk-overhangs into pSB1C3, Cloning ThuricinS with EA silk-overhangs into pSB1C3, Cloning ThuricinS with ED silk-overhangs into pSB1C3 and IMPACT Purification of K2018011

K2018011in standard iGEM plasmid (pSB1C3) was used in a phusion PCR, SOP0010 . The primes have an overhang, which makes the gene compatible with the silk overhang, and thereby enabling us to make a hybrid silk fiber.

pPCR program:

Segment	Step	Temperature	Duration
1	Initial denaturation	98 °C	2 min
2	34 cycles	98 °C	30 sec
		70 °C	30 sec
		72 °C	15 sec
3	Final extension	72 °C	5 min
4	Keep the sample cold	20 °C	HOLD

The result is verified by Gel electrophoresis. The result is shown here:

The lane marked no. 4 and 8 are ThuricinS with ED-overhangs and is expected to be 217 bp long. We have used the yellow ladder, where the fourth band is 200 bp. Based on that it can be assumed that we succeeded at getting silk-overhangs on ThuricinS.

What we had done with the BioBrick K2018011 can be read in the protocols: Cloning composite part K2018011 into pSB1C3, Cloning ThuricinS with DA silk-overhangs into pSB1C3, Cloning ThuricinS with DE silk-overhangs into pSB1C3, Cloning ThuricinS with EA silk-overhangs into pSB1C3, Cloning ThuricinS with ED silk-overhangs into pSB1C3 and IMPACT Purification of K2018011

Week 34

For using the IMPACT method for purification of bacteriocins, some specific restrictions sites are necessary. Therefore a Phusion PCR was made by following SOP0010 .

pPCR program:

Segment	Step	Temperature	Duration
1	Initial denaturation	98 °C	30 sec
2	34 cycles	98 °C	5 sec
		69 °C	20 sec
		72 °C	15 sec
3	Final extension	72 °C	10 min
4	Keep the sample cold	4 °C	HOLD

What we had done with the BioBrickK2018010 can be read in the protocols: Cloning composite part K2018010 into pSB1C3. and IMPACT Purification of K2018010

For using the IMPACT method for purification of bacteriocins, some specific restrictions sites are necessary. Therefore a Phusion PCR was made by following SOP0010 .

pPCR program:

Segment	Step	Temperature	Duration
1	Initial denaturation	98 °C	30 sec
2	34 cycles	98 °C	5 sec
		69 °C	20 sec
		72 °C	15 sec
3	Final extension	72 °C	10 min
4	Keep the sample cold	4 °C	HOLD

What we had done with the BioBrick K2018011 can be read in the protocols: Cloning composite part K2018011 into pSB1C3, Cloning ThuricinS with DA silk-overhangs into pSB1C3, Cloning ThuricinS with DE silk-overhangs into pSB1C3, Cloning ThuricinS with EA silk-overhangs into pSB1C3, Cloning ThuricinS with ED silk-overhangs into pSB1C3 and IMPACT Purification of K2018011

Week 35

The BioBrick was digested with with SpeI for 2h and EcoRI for 30 min, by following SOP0017 .

After digesting, the sample was ligated by following SOP0015 , with the standard iGEM plasmid, pSB1C3, overnight, and thereafter transformed into E. coli Top10, by following SOP0009. The ratio of the ligation is shown hereunder:

Ratio	1:3	1:5	1:10
Ligase Buffer	2 µl	2 µl	2 µl
Ligase	1 µl	1 µl	1µl
11	3 µl	5 µl	10 µl
BR123	1 µl	1 µl	1 µl
ddH2O	13 µl	11 µl	6 µl

cPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	95 °C	2 min
2	34 cycles	95 °C	15 sec
		60 °C	15 sec
		72 °C	15 sec
3	Final extension	72 °C	2.5 min
4	Keep the sample cold	4 °C	HOLD

The samples 11-17 are from colonies containing plasmids with ThuricinS with DA overhang

ThuricinS with silk overhangs, together with primer overhangs (VF2 and VR), is 517 bp long. Fermentas Generuler 1 kb ladder plus was used, so from these pictures it can verified that ThuricinS with DA overhangs have the right length.

What we had done with the BioBrick K2018011 can be read in the protocols: Cloning composite part K2018011 into pSB1C3, Cloning ThuricinS with DA silk-overhangs into pSB1C3, Cloning ThuricinS with DE silk-overhangs into pSB1C3, Cloning ThuricinS with EA silk-overhangs into pSB1C3, Cloning ThuricinS with ED silk-overhangs into pSB1C3 and IMPACT Purification of K2018011

The BioBrick was digested with with SpeI for 2h and EcoRI for 30 min, by following SOP0017 .

After digesting, the sample was ligated by following SOP0015 , with the standard iGEM plasmid, pSB1C3, overnight, and thereafter transformed into E. coli Top10, by following SOP0009. The ratio of the ligation is shown hereunder:

Ratio	1:3	1:5	1:10
Ligase Buffer	2 µl	2 µl	2 µl
Ligase	1 µl	1 µl	1µl
13	3 µl	5 µl	10 µl
BR123	1 µl	1 µl	1 µl
ddH2O	13 µl	11 µl	6 µl

cPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	95 °C	2 min
2	34 cycles	95 °C	15 sec
		60 °C	15 sec
		72 °C	15 sec
3	Final extension	72 °C	2.5 min
4	Keep the sample cold	4 °C	HOLD

The samples 25- 28 are from colonies containing plasmids with ThuricinS with DE overhang

ThuricinS with silk overhangs, together with primer overhangs (VF2 and VR), is 517 bp long. Fermentas Generuler 1 kb ladder plus was used, so from these pictures it can verified that ThuricinS with DE overhangs have the right length.

What we had done with the BioBrick K2018011 can be read in the protocols: Cloning composite part K2018011 into pSB1C3, Cloning ThuricinS with DA silk-overhangs into pSB1C3, Cloning ThuricinS with DE silk-overhangs into pSB1C3, Cloning ThuricinS with EA silk-overhangs into pSB1C3, Cloning ThuricinS with ED silk-overhangs into pSB1C3 and IMPACT Purification of K2018011

The BioBrick was digested with with SpeI for 2h and EcoRI for 30 min, by following SOP0017 .

After digesting, the sample was ligated by following SOP0015 , with the standard iGEM plasmid, pSB1C3, overnight, and thereafter transformed into E. coli Top10, by following SOP0009. The ratio of the ligation is shown hereunder:

Ratio	1:3	1:5	1:10
Ligase Buffer	2 µl	2 µl	2 µl
Ligase	1 µl	1 µl	1µl
12	3 µl	5 µl	10 µl
BR123	1 µl	1 µl	1 µl
ddH2O	13 µl	11 µl	6 µl

cPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	95 °C	2 min
2	34 cycles	95 °C	15 sec
		60 °C	15 sec
		72 °C	15 sec
3	Final extension	72 °C	2.5 min
4	Keep the sample cold	4 °C	HOLD

The samples 18-24 are from colonies containing plasmids with ThuricinS with EA overhang

ThuricinS with silk overhangs, together with primer overhangs (VF2 and VR), is 517 bp long. Fermentas Generuler 1 kb ladder plus was used, so from these pictures it can verified that ThuricinS with EA overhangs have the right length.

What we had done with the BioBrick K2018011 can be read in the protocols: Cloning composite part K2018011 into pSB1C3, Cloning ThuricinS with DA silk-overhangs into pSB1C3, Cloning ThuricinS with DE silk-overhangs into pSB1C3, Cloning ThuricinS with EA silk-overhangs into pSB1C3, Cloning ThuricinS with ED silk-overhangs into pSB1C3 and IMPACT Purification of K2018011

The BioBrick was digested with with SpeI for 2h and EcoRI for 30 min, by following SOP0017 .

After digesting, the sample was ligated by following SOP0015 , with the standard iGEM plasmid, pSB1C3, overnight, and thereafter transformed into E. coli Top10, by following SOP0009. The ratio of the ligation is shown hereunder:

Ratio	1:3	1:5	1:10
Ligase Buffer	2 µl	2 µl	2 µl
Ligase	1 µl	1 µl	1µl
10	3 µl	5 µl	10 µl
BR123	1 µl	1 µl	1 µl
ddH2O	13 µl	11 µl	6 µl

cPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	95 °C	2 min
2	34 cycles	95 °C	15 sec
		60 °C	15 sec
		72 °C	15 sec
3	Final extension	72 °C	2.5 min
4	Keep the sample cold	4 °C	HOLD

The samples 1-10 are from colonies containing plasmids with ThuricinS with ED overhang

ThuricinS with silk overhangs, together with primer overhangs (VF2 and VR), is 517 bp long. Fermentas Generuler 1 kb ladder plus was used, so from these pictures it can verified that ThuricinS with ED overhangs have the right length.

What we had done with the BioBrick K2018011 can be read in the protocols: Cloning composite part K2018011 into pSB1C3, Cloning ThuricinS with DA silk-overhangs into pSB1C3, Cloning ThuricinS with DE silk-overhangs into pSB1C3, Cloning ThuricinS with EA silk-overhangs into pSB1C3, Cloning ThuricinS with ED silk-overhangs into pSB1C3 and IMPACT Purification of K2018011

Week 36

The BioBrick was first digested with restrictions enzymes by following SOP0017 . The restriction enzymes were: PstI and XbaI for 2h. After digesting, the sample was verified by Gel electrophoresis.

pPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	95 °C	3 min
2	34 cycles	95 °C	25 sec
		58 °C	25 sec
		72 °C	30 sec
3	Final extension	72 °C	1 min
4	Keep the sample cold	20 °C	HOLD

In the next step the vector was ligated with the insert overnight by following SOP0015 , and thereafter it was transformed, SOP0009, into E. coli Top10. The ratio of the ligation is shown hereunder:

Ratio	1:2	1:5
Ligase Buffer	2 µl	2 µl
Ligase	1 µl	1 µl
SG74	2 µl	5 µl
SG26	1 µl	1 µl
ddH2O	14 µl	11 µl

To verify that the transformation and ligation succeeded, a colony PCR was run, SOP0021 .

cPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	95 °C	3 min
2	34 cycles	95 °C	25 sec
		58 °C	25 sec
		72 °C	30 sec
3	Final extension	72 °C	1 min
4	Keep the sample cold	20 °C	HOLD

The Gel Photo below shows the cPCR results. VR and VF2 primers adds a total of 300 bp, the gene is 119 bp, which gives a total DNA length of 419 bp. From this gel the ligation of the gen incorporated into psB1C3 can be observed.

This is the picture of the colony PCR where 1.2 to 1.6 are different colonies of the same plate for MaSp1 CD. 1.3 was used to purify plasmids.

The second gel photo shows cutting of SB100 with Xbal and PstI. Every second well had added restriction enzymes. If a band of 145 bp long was observed on the gel of ligation and transformation of SB100, it was concluded successful and sent to sequencing.

What we have done with the BioBrick K2018043 can be read in the protocol: >Cloning Basic part K2018043 into pSB1C3

The BioBrick was first digested with restrictions enzymes by following SOP0017 . The restriction enzymes were: PstI and XbaI for 2 h. After digesting, the sample was verified by Gel electrophoresis.

pPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	95 °C	3 min
2	34 cycles	95 °C	25 sec
		58 °C	25 sec
		72 °C	30 sec
3	Final extension	72 °C	1 min
4	Keep the sample cold	20 °C	HOLD

In the next step the vector was ligated with the insert overnight by following SOP0015 , and thereafter it was transformed, SOP0009, into E. coli Top10. The ratio of the ligation is shown hereunder:

Ratio	1:2	1:5
Ligase Buffer	2 µl	2 µl
Ligase	1 µl	1 µl
SG75	2 µl	5 µl
SG26	1 µl	1 µl
ddH2O	14 µl	11 µl

To verify that the transformation and ligation succeeded, a colony PCR was run, SOP0021 .

cPCR program used:

Segment	Step	Temperature	Duration
1	Initial denaturation	95 °C	3 min
2	34 cycles	95 °C	25 sec
		58 °C	25 sec
		72 °C	30 sec
3	Final extension	72 °C	1 min
4	Keep the sample cold	20 °C	HOLD

The Gel Photo below shows the cPCR results. VR and VF2 primers adds a total of 300 bp, the gene is 119 bp long, which gives a total DNA length of 419 bp. From this gel the ligation of the gen incorporated into psB1C3 can be indicated .

This is the picture of the colony PCR where 2.1 to 2.3 are different colonies of the same plate for MaSp2 CD. 2.1 was used to purify plasmids

The second gel photo shows cutting of SB101 with Xbal and PstI. Every second well had added restriction enzymes. From this gel, the ligation and transformation of SB101, a band around 145 bp after digestion, was concluded as being successful and sent to sequencing.

What we have done with the BioBrick K2018044 can be read in the protocol: Cloning Basic part K2018044 into pSB1C3

Before purifying the bacteriocin by IMPACT, the bacteriocin must first be transformed into the IMPACT vector bacteria, E. Coli:ER2566. To do this, we incorporated the pPCR product into the IMPACT plasmid, pTXB1. Thereafter the products are digested by following the SOP0025. step. 8.1.1. and afterwards ligated by following step 8.1.3 → 8.1.4 in the same SOP.

The following gel image were used to verify that K2018010 were successfully cloned into pTXB1 (the IMPACT vector). Lane 4-6 is K2018010/pTXB1. Lane 4 and 6 showed the correct sized band. Lane 5 is a negative.

What we had done with the BioBrickK2018010 can be read in the protocols: Cloning composite part K2018010 into pSB1C3. and IMPACT Purification of K2018010

Before purifying the bacteriocin by IMPACT, the bacteriocin must first be transformed into the IMPACT vector bacteria, E. Coli:ER2566. To do this, we incorporated the pPCR product into the IMPACT plasmid, pTXB1. Thereafter the products are digested by following the SOP0025. step. 8.1.1. and afterwards ligated by following step 8.1.3 → 8.1.4 in the same SOP.

The following gel image were used to verify that K2018011 were successfully cloned into pTXB1 (the IMPACT vector). Lane 4-6 is K2018010/pTXB1. Lane 1-3 is K2018011/pTXB1 and all showed the correct sized band.

What we had done with the BioBrick K2018011 can be read in the protocols: Cloning composite part K2018011 into pSB1C3, Cloning ThuricinS with DA silk-overhangs into pSB1C3, Cloning ThuricinS with DE silk-overhangs into pSB1C3, Cloning ThuricinS with EA silk-overhangs into pSB1C3, Cloning ThuricinS with ED silk-overhangs into pSB1C3 and IMPACT Purification of K2018011

Before purifying the bacteriocin by IMPACT, the bacteriocin must first be transformed into the IMPACT vector bacteria, E. Coli:ER2566. To do this, we incorporated the pPCR product into the IMPACT plasmid, pTXB1. Thereafter the products are digested by following the SOP0025. step. 8.1.1. and afterwards ligated by following step 8.1.3 → 8.1.4 in the same SOP.

The following gel image were used to verify that K2018012 were successfully cloned into pTXB1 (the IMPACT vector). Lane 9-12 is K2018012/pTXB1 and they all showed the correct sized band.

What we had done with the BioBrick K2018012 can be read in the protocols: Cloning composite part K2018012 into pSB1C3 and IMPACT Purification of K2018012

Before purifying the bacteriocin by IMPACT, the bacteriocin must first be transformed into the IMPACT vector bacteria, E. Coli:ER2566. To do this, we incorporated the pPCR product into the IMPACT plasmid, pTXB1. Thereafter the products are digested by following the SOP0025. step. 8.1.1. and afterwards ligated by following step 8.1.3 → 8.1.4 in the same SOP.

The following gel image were used to verify that K2018014 were successfully cloned into pTXB1 (the IMPACT vector). The three first lanes is K2018014/pTXB1, where lane 1 and 3 showed the correct sized band but lane 2 was negative..

What we had done with the BioBrickK2018014 can be read in the protocols: Cloning composite part K2018014 into pSB1C3 and IMPACT Purification of K2018014

Before purifying the bacteriocin by IMPACT, the bacteriocin must first be transformed into the IMPACT vector bacteria, E. Coli:ER2566. To do this, we incorporated the pPCR product into the IMPACT plasmid, pTXB1. Thereafter the products are digested by following the SOP0025. step. 8.1.1. and afterwards ligated by following step 8.1.3 → 8.1.4 in the same SOP.

The following gel image were used to verify that K2018015 were successfully cloned into pTXB1 (the IMPACT vector). We used three different primers to ‘cross amplify’ the gene and a part of the vector plasmid, getting 2 different DNA bands. All of the given results were, as expected, at 330 bp and 494 bp.

What we had done with the BioBrick K2018015 can be read in the protocols: <Cloning composite part K2018015 into pSB1C3and IMPACT Purification of K2018015

Week 37

Week 38

The IMPACT purification is done by following the SOP0025. step. 8.1.6→ 8.4.3. and the purity is determined by Bradford Protein Concentration determination.

In the Bradford assay, we measured OD595= 0.09400, and together with the BSA standard curve (see appendix) we calculated the final protein concentration as being 67.1 µg/ml.

What we had done with the BioBrickK2018010 can be read in the protocols: Cloning composite part K2018010 into pSB1C3. and IMPACT Purification of K2018010

The IMPACT purification is done by following the SOP0025. step. 8.1.6→ 8.4.3. and the purity is determined by Bradford Protein Concentration determination.

In the Bradford assay we measured OD595= 0.05700, and together with the BSA standard curve (see appendix) we calculated a final protein concentration as being 33.5 µg/ml.

What we had done with the BioBrick K2018011 can be read in the protocols: Cloning composite part K2018011 into pSB1C3, Cloning ThuricinS with DA silk-overhangs into pSB1C3, Cloning ThuricinS with DE silk-overhangs into pSB1C3, Cloning ThuricinS with EA silk-overhangs into pSB1C3, Cloning ThuricinS with ED silk-overhangs into pSB1C3 and IMPACT Purification of K2018011

The IMPACT purification is done by following the SOP0025. step. 8.1.6→ 8.4.3. and the purity is determined by Bradford Protein Concentration determination.

In the Bradford assy, we measured an OD595= 0.06900, and together with the BSA standard curve (see appendix) we calculated a final protein concentration as being 44.4 µg/ml.

What we had done with the BioBrick K2018012 can be read in the protocols: Cloning composite part K2018012 into pSB1C3 and IMPACT Purification of K2018012

The IMPACT purification is done by following the SOP0025. step. 8.1.6→ 8.4.3. and the purity is determined by Bradford Protein Concentration determination.

In the Bradford assay we measured an OD595= 0.05360, and together with the BSA standard curve (see appendix) we calculated a final protein concentration as being 30.4 µg/ml.

What we had done with the BioBrickK2018014 can be read in the protocols: Cloning composite part K2018014 into pSB1C3 and IMPACT Purification of K2018014

The IMPACT purification is done by following the SOP0025. step. 8.1.6→ 8.4.3. and the purity is determined by Bradford Protein Concentration determination.

In the Bradford assay we measured OD595= 0.07800, and together with the BSA standard curve (see appendix) we calculated a final protein concentration as being 52.5 µg/ml.

What we had done with the BioBrick K2018015 can be read in the protocols: <Cloning composite part K2018015 into pSB1C3and IMPACT Purification of K2018015

Week 39

The effect of the bacteriocins is validated by a MIC-test, which is made by following the SOP0027 step. 7.2→ 7.9.

The MIC assay gave the following MIC values:

Strain	MIC (µg/ml)
S. aureus:CC398	33.6
S. aureus:USA300	16.8
S. aureus:hVISA	16.8
P. aeruginosa:PAO1	33.6

What we had done with the BioBrickK2018010 can be read in the protocols: Cloning composite part K2018010 into pSB1C3. and IMPACT Purification of K2018010

The effect of the bacteriocins is validated by a MIC-test, which is made by following the SOP0027 step. 7.2→ 7.9.

The MIC assay gave the following MIC values:

Strain	MIC (µg/ml)
S. aureus:CC398	16.7
S. aureus:USA300	16.7
S. aureus:hVISA	16.7
P. aeruginosa:PAO1	16.7

What we had done with the BioBrick K2018011 can be read in the protocols: Cloning composite part K2018011 into pSB1C3, Cloning ThuricinS with DA silk-overhangs into pSB1C3, Cloning ThuricinS with DE silk-overhangs into pSB1C3, Cloning ThuricinS with EA silk-overhangs into pSB1C3, Cloning ThuricinS with ED silk-overhangs into pSB1C3 and IMPACT Purification of K2018011

The effect of the bacteriocins is validated by a MIC-test, which is made by following the SOP0027 step. 7.2→ 7.9.

The MIC assay gave the following MIC values:

Strain	MIC (µg/ml)
S. aureus:CC398	22.2
S. aureus:USA300	22.2
S. aureus:hVISA	22.2
P. aeruginosa:PAO1	>22.2

What we had done with the BioBrick K2018012 can be read in the protocols: Cloning composite part K2018012 into pSB1C3 and IMPACT Purification of K2018012

The effect of the bacteriocins is validated by a MIC-test, which is made by following the SOP0027 step. 7.2→ 7.9.

The MIC assay gave the following MIC values:

Strain	MIC (µg/ml)
S. aureus:CC398	13.1
S. aureus:USA300	12.1
S. aureus:hVISA	6.6
P. aeruginosa:PAO1	>13.1

What we had done with the BioBrickK2018014 can be read in the protocols: Cloning composite part K2018014 into pSB1C3 and IMPACT Purification of K2018014

The effect of the bacteriocins is validated by a MIC-test, which is made by following the SOP0027 step. 7.2→ 7.9.

The MIC assay gave the following MIC values:

Strain	MIC (µg/ml)
S. aureus:CC398	7.6
S. aureus:USA300	7.6
S. aureus:hVISA	7.6
P. aeruginosa:PAO1	>15.2

What we had done with the BioBrick K2018015 can be read in the protocols: <Cloning composite part K2018015 into pSB1C3and IMPACT Purification of K2018015

@@ Line 16: / Line 16: @@
 <!-------------------------------------------Week 10 ------------------------------------------------------>
 <h4><b>Week 10</b></h4>
-<button class="accordion"><img src=" https://static.igem.org/mediawiki/2016/5/55/T--SDU-Denmark--socialmini.png" height="20%" style="float:right;margin-left:15px;"> The team meets for the first time. Date 16.03.09</button>
+<button class="accordion"><img src=" https://static.igem.org/mediawiki/2016/5/55/T--SDU-Denmark--socialmini.png" width="70%" style="margin:auto;"> The team meets for the first time. Date 16.03.09</button>
 <div id="foo" class="panel">
 </div>
-<button class="accordion"><img src=" https://static.igem.org/mediawiki/2016/5/55/T--SDU-Denmark--socialmini.png" height="20%" style="float:right;margin-left:15px;"> Idea generation weekend with the team, in a weekend cottage fare, fare away from the university. Date 16.03.11 to 16.03.13
+<button class="accordion"><img src=" https://static.igem.org/mediawiki/2016/5/55/T--SDU-Denmark--socialmini.png" width="70%" style="margin:auto;"> Idea generation weekend with the team, in a weekend cottage fare, fare away from the university. Date 16.03.11 to 16.03.13
 </button>
 <div id="foo" class="panel">
-<p><img src="https://static.igem.org/mediawiki/2016/1/14/T--SDU-Denmark--social_1.png" width=”70%” style=”margin:auto;”/></p>
+<p><img src="https://static.igem.org/mediawiki/2016/1/14/T--SDU-Denmark--social_1.png" width="70%" style="margin:auto;"></p>
-<p><img src="https://static.igem.org/mediawiki/2016/a/ab/T--SDU-Denmark--social_3.jpg" " width=”70%” style=”margin:auto;”/></p>
+<p><img src="https://static.igem.org/mediawiki/2016/a/ab/T--SDU-Denmark--social_3.jpg" width="70%" style="margin:auto;"></p>
 </div>
 <!-------------------------------------------Week 13 ------------------------------------------------------>
 <h4><b>Week 13</b></h4>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/5/55/T--SDU-Denmark--socialmini.png" height="20%" style="float:right;margin-left:15px;"> The project is choosed. Date 16.03.28 </button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/5/55/T--SDU-Denmark--socialmini.png" width="70%" style="margin:auto;"> The project is choosed. Date 16.03.28 </button>
 <div id="foo" class="panel">
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/5/55/T--SDU-Denmark--socialmini.png" height="20%" style="float:right;margin-left:15px;"> The first project description is submitted to the advisors. 16.04.01</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/5/55/T--SDU-Denmark--socialmini.png" width="70%" style="margin:auto;"> The first project description is submitted to the advisors. 16.04.01</button>
 <div id="foo" class="panel">
 </div>
 <!-------------------------------------------Week 14 ------------------------------------------------------>
 <h4><b>Week 14</b></h4>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/5/55/T--SDU-Denmark--socialmini.png" height="20%" style="float:right;margin-left:15px;"> The project is present for the advisors. Date 16.04.04</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/5/55/T--SDU-Denmark--socialmini.png" width="70%" style="margin:auto;"> The project is present for the advisors. Date 16.04.04</button>
 <div id="foo" class="panel">
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/5/55/T--SDU-Denmark--socialmini.png" height="20%" style="float:right;margin-left:15px;"> We held a presentation with BioOL. Date 16.04.07</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/5/55/T--SDU-Denmark--socialmini.png" width="70%" style="margin:auto;"> We held a presentation with BioOL. Date 16.04.07</button>
 <div id="foo" class="panel">
 </div>
 <!-------------------------------------------Week 15 ------------------------------------------------------>
 <h4><b>Week 15</b></h4>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/5/55/T--SDU-Denmark--socialmini.png" height="20%" style="float:right;margin-left:15px;"> Laboratory course Date 16.04.14-16.04.16</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/5/55/T--SDU-Denmark--socialmini.png" width="70%" style="margin:auto;"> Laboratory course Date 16.04.14-16.04.16</button>
 <div id="foo" class="panel">
-<p> We leaned why we find thing in the lab, and try to make some basic experiments. On the photo is Joel learning to pipet</p>
+<p> We learned how we find thing in the lab, and tried to make some basic experiments. On the photo is Joel learning how to pipet</p>
-<p><img src="https://static.igem.org/mediawiki/2016/4/4a/T--SDU-Denmark--social_6.jpg" " width=”70%” style=”margin:auto;”/></p>
+<p><img src="https://static.igem.org/mediawiki/2016/4/4a/T--SDU-Denmark--social_6.jpg" width=”70%” style=”margin:auto;”></p>
 </div>
 <!-------------------------------------------Week 17 ------------------------------------------------------>
 <h4><b>Week 17</b></h4>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/5/55/T--SDU-Denmark--socialmini.png" height="20%" style="float:right;margin-left:15px;"> Research day at the university. Date 16.04.30</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/5/55/T--SDU-Denmark--socialmini.png" width="70%" style="margin:auto;"> Research day at the university. Date 16.04.30</button>
 <div id="foo" class="panel">
 <p> This year the faculty of Biochemistry and Molecular Biology had a big workshop area with the theme <a target="blank" href="http://www.sdu.dk/forskning/forskningsformidling/forskningensdoegn/odense/for_hele_familien/den_gode_onde_grusomme">“The Good, the Bad and the Ugly”</a>: the probiotic bacteria, the pathogenic bacteria and the multiresistant bacteria.</p>
-<p><img src="https://static.igem.org/mediawiki/2016/6/67/T--SDU-Denmark--social_7.jpg" " width=”70%” style=”margin:auto;”/></p>
+<p><img src="https://static.igem.org/mediawiki/2016/6/67/T--SDU-Denmark--social_7.jpg" width="70%" style="margin:auto;"></p>
 </div>
 <!-------------------------------------------Week 18 ------------------------------------------------------>
 <h4><b>Week 18</b></h4>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/5/55/T--SDU-Denmark--socialmini.png" height="20%" style="float:right;margin-left:15px;"> Danish Meetup at Danish Technical University. Date 16.05.06-16.05-08</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/5/55/T--SDU-Denmark--socialmini.png" width="70%" style="margin:auto;"> Danish Meetup at Danish Technical University. Date 16.05.06-16.05-08</button>
 <div id="foo" class="panel">
-<p><img src="https://static.igem.org/mediawiki/2016/8/80/T--SDU-Denmark--social_8.jpg" " width=”70%” style=”margin:auto;” /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/8/80/T--SDU-Denmark--social_8.jpg" width="70%" style="margin:auto;"></p>
-<p><img src="https://static.igem.org/mediawiki/2016/8/89/T--SDU-Denmark--social_9.jpg" " width=”70%” style=”margin:auto;”/></p>
+<p><img src="https://static.igem.org/mediawiki/2016/8/89/T--SDU-Denmark--social_9.jpg" width="70%" style="margin:auto;"></p>
 </div>
 <!-------------------------------------------Week 24 ------------------------------------------------------>
@@ Line 67: / Line 67: @@
 <button class="accordion"><img src="https://static.igem.org/mediawiki/2016/5/55/T--SDU-Denmark--socialmini.png" height="20%" style="float:right;margin-left:15px;"> Nordic Meetup in Stockholm. Date 16.06.16-16.06-19</button>
 <div id="foo" class="panel">
-<p><img src="https://static.igem.org/mediawiki/2016/1/16/T--SDU-Denmark--social_10.jpg" " width=”70%” style=”margin:auto;”/></p>
+<p><img src="https://static.igem.org/mediawiki/2016/1/16/T--SDU-Denmark--social_10.jpg" width="70%" style="margin:auto;"></p>
-<p><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--social_11.jpg" " width=”70%” style=”margin:auto;”/></p>
+<p><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--social_11.jpg" width="70%" style="margin:auto;"></p>
 </div>
 <!-------------------------------------------Week 26 ------------------------------------------------------>
 <h4><b>Week 26</b></h4>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" height="20%" style="float:right;margin-left:15px;">Received gBlock, K201843, and resuspend. Date: 16.06.27</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" width="70%" style="margin:auto;">Received gBlock, K201843, and resuspend. Date: 16.06.27</button>
 <div id="foo" class="panel">
 <p>The brick was ordered from IDT and received: The received gblock had a weight of 200 ng. and was resuspended in dH2O.
@@ Line 82: / Line 82: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" height="20%" style="float:right;margin-left:15px;">Received gBlock, K201844, and resuspend. Date: 16.06.27</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" width="70%" style="margin:auto;">Received gBlock, K201844, and resuspend. Date: 16.06.27</button>
 <div id="foo" class="panel">
 <p>The brick was ordered from IDT and received: The received gblock had a weight of 200 ng. and was resuspended in dH2O.
@@ Line 91: / Line 91: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" height="20%" style="float:right;margin-left:15px;">Received gBlock, K1763002, and resuspended. Date: 16.06.27</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" width="70%" style="margin:auto;">Received gBlock, K1763002, and resuspended. Date: 16.06.27</button>
 <div id="foo" class="panel">
 <p>The brick was ordered from IDT and received: The received gblock had a weight of 200 ng. and was resuspended in dH2O.
@@ Line 100: / Line 100: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" height="20%" style="float:right;margin-left:15px;">Received gBlock, K1763003, and resuspended. Date: 16.06.27</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" width="70%" style="margin:auto;">Received gBlock, K1763003, and resuspended. Date: 16.06.27</button>
 <div id="foo" class="panel">
 <p>The brick was ordered from IDT and received: The received gblock had a weight of 200 ng. and was resuspended in dH2O.
@@ Line 109: / Line 109: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" height="20%" style="float:right;margin-left:15px;">Received gBlock, K1763004, and resuspended. Date: 16.06.27</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" width="70%" style="margin:auto;">Received gBlock, K1763004, and resuspended. Date: 16.06.27</button>
 <div id="foo" class="panel">
 <p>The brick was ordered from IDT and received: The received gblock had a weight of 200 ng. and was resuspended in dH2O.
@@ Line 118: / Line 118: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" height="20%" style="float:right;margin-left:15px;">Received gBlock, K1763009, and resuspended. Date: 16.06.27</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" width="70%" style="margin:auto;">Received gBlock, K1763009, and resuspended. Date: 16.06.27</button>
 <div id="foo" class="panel">
 <p>The brick was ordered from IDT and received: The received gblock had a weight of 200 ng. and was resuspended in dH2O.
@@ Line 127: / Line 127: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" height="20%" style="float:right;margin-left:15px;">Received gBlock, K1763010, and resuspended. Date: 16.06.27 </button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" width="70%" style="margin:auto;">Received gBlock, K1763010, and resuspended. Date: 16.06.27 </button>
 <div id="foo" class="panel">
 <p>The brick was ordered from IDT and received: The received gblock had a weight of 200 ng. and was resuspended in dH2O.
@@ Line 135: / Line 135: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" height="20%" style="float:right;margin-left:15px;">Received gBlock, K1763011, and resuspended. Date: 16.06.27 </button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" width="70%" style="margin:auto;">Received gBlock, K1763011, and resuspended. Date: 16.06.27 </button>
 <div id="foo" class="panel">
 <p>The brick was ordered from IDT and received: The received gblock had a weight of 200 ng. and was resuspended in dH2O.
@@ Line 143: / Line 143: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" height="20%" style="float:right;margin-left:15px;">Received gBlock, K1763012, and resuspended. Date: 16.06.27</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" width="70%" style="margin:auto;">Received gBlock, K1763012, and resuspended. Date: 16.06.27</button>
 <div id="foo" class="panel">
 <p>The brick was ordered from IDT and received: The received gblock had a weight of 200 ng. and was resuspended in dH2O.
@@ Line 155: / Line 155: @@
 <!-------------------------------------------Week 27 ------------------------------------------------------>
 <h4><b>Week 27</b></h4>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Received plasmid, K2018011, and transformation into Top10 <em>E. coli</em>. Date: 16.07.05</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Received plasmid, K2018011, and transformation into Top10 <em>E. coli</em>. Date: 16.07.05</button>
 <div id="foo" class="panel">
 <p>The BioBrick was ordered from IDT in a plasmid pUCIDT-AMP and after receiving it, it was resuspended to a final concentration of 0.1 µg/µl. </p>
@@ Line 168: / Line 168: @@
 <!-------------------------------------------Week 28 ------------------------------------------------------>
 <h4><b>Week 28</b></h4>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Received plasmid, K2018010, and transformation into Top10 <em>E. coli</em>. Date: 16.07.14</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Received plasmid, K2018010, and transformation into Top10 <em>E. coli</em>. Date: 16.07.14</button>
 <div id="foo" class="panel">
 <p>The BioBrick was ordered from IDT in a plasmid pUCIDT-AMP and after receiving it, it was resuspended to a final concentration of 0.1 µg/µl.</p>
@@ Line 179: / Line 179: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Verification of K2018011 in standard iGEM plasmid (pSB1C3). Date: 16.07.14</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Verification of K2018011 in standard iGEM plasmid (pSB1C3). Date: 16.07.14</button>
 <div id="foo" class="panel">
 <p>The BioBrick containing the gene  <a href="http://parts.igem.org/Part:BBa_K2018011" target="_blank">K2018011</a> was digested with PstI and XbaI overnight by using <a href=" https://static.igem.org/mediawiki/2016/b/b8/T--SDU-Denmark--SOP0017-FastDigest.pdf" target="_blank"> SOP0017 </a>.<p>
@@ Line 234: / Line 234: @@
 <p>After a colony PCR, <a href="https://static.igem.org/mediawiki/2016/f/f9/T--SDU-DENMARK--SOP0021-ColonyPCRwithMyTag.pdf" target="_blank">SOP0021 </a>, was run and verified on a gel, the following result was given:
 Our part is 259 bp and primer overhangs are 150 X 2 bp, which is, in total, 559 bp: A gel electrophoresis from cPCR showed a band around 600 bp, so we assumed that we had the correct  part inserted into the iGEM standard plasmid.</p>
-<p><img src="https://static.igem.org/mediawiki/2016/3/33/T--SDU-Denmark--Notebook--1_and_.png" alt="" width="317" height="239" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/3/33/T--SDU-Denmark--Notebook--1_and_.png" width="70%" style="margin:auto;"></p>
 <p>What we had done with the BioBrick <a href="http://parts.igem.org/Part:BBa_K2018011" target="_blank">K2018011</a> can be read in the protocols: <a href="https://static.igem.org/mediawiki/2016/e/ea/T--SDU-Denmark--K2018011clonedintopSB1C3.pdf" target="_blank">Cloning composite part K2018011 into pSB1C3</a>,  <a href="https://static.igem.org/mediawiki/2016/1/12/T--SDU-Denmark--CloningThuricinwithDAsilk-overhangsintopSB1C3.pdf" target="_blank">Cloning ThuricinS with DA silk-overhangs into pSB1C3</a>, <a href="https://static.igem.org/mediawiki/2016/0/0d/T--SDU-Denmark--CloningThuricinwithDEsilk-overhangsintopSB1C3.pdf" target="_blank">Cloning ThuricinS with DE silk-overhangs into pSB1C3</a>, <a href="https://static.igem.org/mediawiki/2016/5/52/T--SDU-Denmark--CloningThuricinwithEAsilk-overhangsintopSB1C3.pdf" target="_blank">Cloning ThuricinS with EA silk-overhangs into pSB1C3</a>, <a href="https://static.igem.org/mediawiki/2016/b/b1/T--SDU-Denmark--CloningThuricinwithEDsilk-overhangsintopSB1C3.pdf" target="_blank">Cloning ThuricinS with ED silk-overhangs into pSB1C3</a> and <a href="https://static.igem.org/mediawiki/2016/0/03/T--SDU-Denmark--IMPACTPurificationofK2018011.pdf" target="_blank">IMPACT Purification of K2018011</a> </p>
@@ Line 241: / Line 241: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Received plasmid, K2018014, and transformation into Top10 <em>E. coli</em>. Date: 16.07.14</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Received plasmid, K2018014, and transformation into Top10 <em>E. coli</em>. Date: 16.07.14</button>
 <div id="foo" class="panel">
 <p>The BioBrick was ordered from IDT in a plasmid pUCIDT-AMP and after receiving it, it was resuspended to a final concentration of 0.1 µg/µl.</p>
@@ Line 252: / Line 252: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Received plasmid, K2018019, and transformation into Top10 <em>E. coli</em>. Date: 16.07.14</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Received plasmid, K2018019, and transformation into Top10 <em>E. coli</em>. Date: 16.07.14</button>
 <div id="foo" class="panel">
 <p>The BioBrick was ordered from IDT in a plasmid pUCIDT-AMP and after receiving it, it was resuspended to a final concentration of 0.1 µg/µl. </p>
@@ Line 267: / Line 267: @@
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" height="20%" style="float:right;margin-left:15px;">Verification of K1763002, in standard iGEM plasmid (pSB1C3). Date: 16.07.24</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" width="70%" style="margin:auto;">Verification of K1763002, in standard iGEM plasmid (pSB1C3). Date: 16.07.24</button>
 <div id="foo" class="panel">
 <p>The BioBrick was first amplified by phusion PCR, <a href="https://static.igem.org/mediawiki/2016/3/37/T--SDU-Denmark--SOP0010-PhusionPCR.pdf ”target="_blank"> SOP0010 </a>, thereafter digested with restrictions enzymes, following <a href=" https://static.igem.org/mediawiki/2016/b/b8/T--SDU-Denmark--SOP0017-FastDigest.pdf" target="_blank"> SOP0017 </a>. The restriction enzymes were: PstI for 2 h and EcoRI for 30 min. After digesting, the sample was verified by running a gel electrophoresis. </p>
@@ Line 407: / Line 407: @@
 <p>The Gel Photo below shows the cPCR results. VR and VF2 primers adds a total of 300 bp, the gene is 119 bp which gives a total of DNA length of 419 bp.  From this gel, the ligation of the gen incorporated into psB1C3 can be observed .</p>
-<p><img src="https://static.igem.org/mediawiki/2016/b/b6/T--SDU-Denmark--Notebook--2_and_.png" alt="" width="70%"  /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/b/b6/T--SDU-Denmark--Notebook--2_and_.png" width="70%" style="margin:auto;"></p>
 <p>Picture of cPCR of different colonies on the same plate (each gene had its own plate). All the colony PCR shows a band near the 4-500 bp band, which was desired.</p>
@@ Line 413: / Line 413: @@
 <p>The second gel photo shows cutting of SB71 with Pst1 at 2 hours with 37 &deg;C and EcoR1 at 30 minutes. Every second well had added restriction enzymes.If a band of approximately 160 bp could be seen on the gel, it was concluded as being successful and sent to sequencing.</p>
-<p><img src="https://static.igem.org/mediawiki/2016/6/63/T--SDU-Denmark--Notebook--3_and_.png" alt="" width="461" height="363" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/6/63/T--SDU-Denmark--Notebook--3_and_.png" width="70%" style="margin:auto;"></p>
@@ Line 421: / Line 421: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" height="20%" style="float:right;margin-left:15px;">Verification of K1763003, in standard iGEM plasmid (pSB1C3). Date: 16.07.24</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" width="70%" style="margin:auto;">Verification of K1763003, in standard iGEM plasmid (pSB1C3). Date: 16.07.24</button>
 <div id="foo" class="panel">
 <p>The BioBrick was first amplified by phusion PCR, <a href="https://static.igem.org/mediawiki/2016/3/37/T--SDU-Denmark--SOP0010-PhusionPCR.pdf ”target="_blank"> SOP0010 </a>, and thereafter digested with restrictions enzymes, following <a href=" https://static.igem.org/mediawiki/2016/b/b8/T--SDU-Denmark--SOP0017-FastDigest.pdf" target="_blank"> SOP0017 </a>. The restriction enzymes were: PstI for 2 h and EcoRI for 30 min. After digesting, the sample was verified by running a gel electrophoresis.</p>
@@ Line 561: / Line 561: @@
 <p>The Gel Photo below shows the cPCR results. VR and VF2 primers adds a total of 300 bp, the gene is 119 bp which gives a total of DNA length of 419 bp.  From this gel, the ligation of the gen incorporated into psB1C3 can be observed .</p>
-<p><img src="https://static.igem.org/mediawiki/2016/b/b6/T--SDU-Denmark--Notebook--2_and_.png" alt="" width="557" height="234" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/b/b6/T--SDU-Denmark--Notebook--2_and_.png" width="70%" style="margin:auto;"></p>
 <p>Picture of cPCR of different colonies on the same plate (each gene had its own plate). All the colony PCR shows a band near the 4-500 bp band.</p>
@@ Line 567: / Line 567: @@
 <p>The second gel photo shows cutting of SB73 with Pst1 at 2 hours with 37 &deg;C and EcoR1 at 30 minutes. Every second well had added restriction enzymes.If a band of approximately 160 bp could be seen on the gel, it was concluded as being successful and sent to sequencing.</p>
-<p><img src="https://static.igem.org/mediawiki/2016/6/63/T--SDU-Denmark--Notebook--3_and_.png" alt="" width="461" height="363" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/6/63/T--SDU-Denmark--Notebook--3_and_.png" width="70%" style="margin:auto;"></p>
@@ Line 575: / Line 575: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" height="20%" style="float:right;margin-left:15px;">Verification of K1763004, in standard iGEM plasmid (pSB1C3). Date: 16.07.24</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" width="70%" style="margin:auto;">Verification of K1763004, in standard iGEM plasmid (pSB1C3). Date: 16.07.24</button>
 <div id="foo" class="panel">
 <p>The BioBrick was first amplified by phusion PCR, <a href="https://static.igem.org/mediawiki/2016/3/37/T--SDU-Denmark--SOP0010-PhusionPCR.pdf ”target="_blank"> SOP0010 </a>, and thereafter digested with restrictions enzymes, following <a href=" https://static.igem.org/mediawiki/2016/b/b8/T--SDU-Denmark--SOP0017-FastDigest.pdf" target="_blank"> SOP0017 </a>. The restriction enzymes were: PstI for 2 h and EcoRI for 30 min. After digesting, the sample was verified by running a gel electrophoresis.</p>
@@ Line 715: / Line 715: @@
 <p>The Gel Photo below shows the cPCR results. VR and VF2 primers adds a total of 300 bp, the gene is 119 bp which gives a total of DNA length of 419 bp.  From this gel, the ligation of the gen incorporated into psB1C3 can be seen.</p>
-<p><img src="https://static.igem.org/mediawiki/2016/b/b6/T--SDU-Denmark--Notebook--2_and_.png" alt="" width="557" height="234" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/b/b6/T--SDU-Denmark--Notebook--2_and_.png" width="70%" style="margin:auto;"></p>
 <p>Picture of cPCR of different colonies on the same plate (each gene had its own plate). All the colony PCR shows a band near the 4-500 bp band.</p>
@@ Line 721: / Line 721: @@
 <p>The second gel photo shows cutting of SB75 with Pst1 at 2 hours with 37 &deg;C and EcoR1 at 30 minutes. Every second well had added restriction enzymes.If a band of approximately 160 bp could be seen on the gel, it was concluded as being successful and sent to sequencing.</p>
-<p><img src="https://static.igem.org/mediawiki/2016/6/63/T--SDU-Denmark--Notebook--3_and_.png" alt="" width="461" height="363" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/6/63/T--SDU-Denmark--Notebook--3_and_.png" width="70%" style="margin:auto;"></p>
@@ Line 729: / Line 729: @@
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" height="20%" style="float:right;margin-left:15px;">Verification of K1763009, in standard iGEM plasmid (pSB1C3). Date: 16.07.24</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" width="70%" style="margin:auto;">Verification of K1763009, in standard iGEM plasmid (pSB1C3). Date: 16.07.24</button>
 <div id="foo" class="panel">
 <p>The BioBrick was first amplified by phusion PCR, <a href="https://static.igem.org/mediawiki/2016/3/37/T--SDU-Denmark--SOP0010-PhusionPCR.pdf ”target="_blank"> SOP0010 </a>, and thereafter digested with restrictions enzymes, following <a href=" https://static.igem.org/mediawiki/2016/b/b8/T--SDU-Denmark--SOP0017-FastDigest.pdf" target="_blank"> SOP0017 </a>. The restriction enzymes were: PstI for 2 h and EcoRI for 30 min. After digesting, the sample was verified by running a gel electrophoresis.</p>
@@ Line 869: / Line 869: @@
 <p>The Gel Photo below shows the cPCR results. VR and VF2 primers adds a total of 300 bp, the gene is 119 bp which gives a total of DNA length of 419 bp.  From this gel, the ligation of the gen incorporated into psB1C3 can be observed .</p>
-<p><img src="https://static.igem.org/mediawiki/2016/b/b6/T--SDU-Denmark--Notebook--2_and_.png" alt="" width="557" height="234" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/b/b6/T--SDU-Denmark--Notebook--2_and_.png" width="70%" style="margin:auto;"></p>
 <p>Picture of cPCR of different colonies on the same plate (each gene had its own plate). All the colony PCR shows a band near the 4-500 bp band.</p>
@@ Line 875: / Line 875: @@
 <p>The second gel photo shows cutting of SB77 with Pst1 at 2 hours with 37 &deg;C and EcoR1 at 30 minutes. Every second well had added restriction enzymes.If a band of approximately 160 bp could be seen on the gel, it was concluded as being successful and sent to sequencing.</p>
-<p><img src="https://static.igem.org/mediawiki/2016/6/63/T--SDU-Denmark--Notebook--3_and_.png" alt="" width="461" height="363" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/6/63/T--SDU-Denmark--Notebook--3_and_.png" width="70%" style="margin:auto;"></p>
@@ Line 883: / Line 883: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" height="20%" style="float:right;margin-left:15px;">Verification of K1763010, in standard iGEM plasmid (pSB1C3). Date: 16.07.24</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" width="70%" style="margin:auto;">Verification of K1763010, in standard iGEM plasmid (pSB1C3). Date: 16.07.24</button>
 <div id="foo" class="panel">
 <p>The BioBrick was first amplified by phusion PCR, <a href="https://static.igem.org/mediawiki/2016/3/37/T--SDU-Denmark--SOP0010-PhusionPCR.pdf ”target="_blank"> SOP0010 </a>, and thereafter digested with restrictions enzymes, following <a href=" https://static.igem.org/mediawiki/2016/b/b8/T--SDU-Denmark--SOP0017-FastDigest.pdf" target="_blank"> SOP0017 </a>. The restriction enzymes were: PstI for 2 h and EcoRI for 30 min. After digesting, the sample was verified by running a gel electrophoresis.</p>
@@ Line 1,023: / Line 1,023: @@
 <p>The Gel Photo below shows the cPCR results. VR and VF2 primers adds a total of 300 bp, the gene is 119 bp which gives a total of DNA length of 419 bp.  From this gel, the ligation of the gen incorporated into psB1C3 can be observed.</p>
-<p><img src="https://static.igem.org/mediawiki/2016/6/6c/T--SDU-Denmark--Notebook--4_and_.png" alt="" width="485" height="159" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/6/6c/T--SDU-Denmark--Notebook--4_and_.png" width="70%" style="margin:auto;"></p>
 <p>Picture of cPCR of different colonies on the same plate (each gene had its own plate). All the colony PCR shows a band near the 4-500 bp band.</p>
@@ Line 1,029: / Line 1,029: @@
 <p>The second gel photo shows cutting of SB63 with Pst1 at 2 hours with 37 &deg;C and EcoR1 at 30 minutes. Every second well had added restriction enzymes.If a band of approximately 160 bp could be seen on the gel, it was concluded as being successful and sent to sequencing.</p>
-<p><img src="https://static.igem.org/mediawiki/2016/6/63/T--SDU-Denmark--Notebook--3_and_.png" alt="" width="461" height="363" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/6/63/T--SDU-Denmark--Notebook--3_and_.png" awidth="70%" style="margin:auto;"></p>
@@ Line 1,038: / Line 1,038: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" height="20%" style="float:right;margin-left:15px;">Verification of K1763011, in standard iGEM plasmid (pSB1C3). Date: 16.07.24</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" width="70%" style="margin:auto;">Verification of K1763011, in standard iGEM plasmid (pSB1C3). Date: 16.07.24</button>
 <div id="foo" class="panel">
 <p>The BioBrick was first amplified by phusion PCR, <a href="https://static.igem.org/mediawiki/2016/3/37/T--SDU-Denmark--SOP0010-PhusionPCR.pdf ”target="_blank"> SOP0010 </a>, and thereafter digested with restrictions enzymes, following <a href=" https://static.igem.org/mediawiki/2016/b/b8/T--SDU-Denmark--SOP0017-FastDigest.pdf" target="_blank"> SOP0017 </a>. The restriction enzymes were: PstI for 2 h and EcoRI for 30 min. After digesting, the sample was verified by running a gel electrophoresis.</p>
@@ Line 1,178: / Line 1,178: @@
 <p>The Gel Photo below shows the cPCR results. VR and VF2 primers adds a total of 300 bp, the gene is 119 bp which gives a total of DNA length of 419 bp.  From this gel, the ligation of the gen incorporated into psB1C3 can be observed .</p>
-<p><img src="https://static.igem.org/mediawiki/2016/6/6c/T--SDU-Denmark--Notebook--4_and_.png" alt="" width="485" height="159" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/6/6c/T--SDU-Denmark--Notebook--4_and_.png" width="70%" style="margin:auto;"></p>
 <p>Picture of cPCR of different colonies on the same plate (each gene had its own plate). All the colony PCR shows a band near the 4-500 bp band.</p>
@@ Line 1,184: / Line 1,184: @@
 <p>The second gel photo shows cutting of SB65 with Pst1 at 2 hours with 37 &deg;C and EcoR1 at 30 minutes. Every second well had added restriction enzymes.If a band of approximately 160 bp could be seen on the gel, it was concluded as being successful and sent to sequencing.</p>
-<p><img src="https://static.igem.org/mediawiki/2016/6/63/T--SDU-Denmark--Notebook--3_and_.png" alt="" width="461" height="363" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/6/63/T--SDU-Denmark--Notebook--3_and_.png" width="70%" style="margin:auto;"></p>
@@ Line 1,191: / Line 1,191: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" height="20%" style="float:right;margin-left:15px;">Verification of K1763012, in standard iGEM plasmid (pSB1C3). Date: 16.07.24</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" width="70%" style="margin:auto;">Verification of K1763012, in standard iGEM plasmid (pSB1C3). Date: 16.07.24</button>
 <div id="foo" class="panel">
 <p>The BioBrick was first amplified by phusion PCR, <a href="https://static.igem.org/mediawiki/2016/3/37/T--SDU-Denmark--SOP0010-PhusionPCR.pdf ”target="_blank"> SOP0010 </a>, and thereafter digested with restrictions enzymes, following <a href=" https://static.igem.org/mediawiki/2016/b/b8/T--SDU-Denmark--SOP0017-FastDigest.pdf" target="_blank"> SOP0017 </a>. The restriction enzymes were: PstI for 2 h and EcoRI for 30 min. After digesting, the sample was verified by running a gel electrophoresis.</p>
@@ Line 1,331: / Line 1,331: @@
 <p>The Gel Photo below shows the cPCR results. VR and VF2 primers adds a total of 300 bp, the gene is 119 bp which gives a total of DNA length of 419 bp.  From this gel, the ligation of the gen incorporated into psB1C3 can be observed .</p>
-<p><img src="https://static.igem.org/mediawiki/2016/6/6c/T--SDU-Denmark--Notebook--4_and_.png" alt="" width="485" height="159" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/6/6c/T--SDU-Denmark--Notebook--4_and_.png" width="70%" style="margin:auto;"></p>
 <p>Picture of cPCR of different colonies on the same plate (each gene had its own plate). All the colony PCR shows a band near the 4-500 bp band.</p>
@@ Line 1,337: / Line 1,337: @@
 <p>The second gel photo shows cutting of SB67 with Pst1 at 2 hours with 37 &deg;C and EcoR1 at 30 minutes. Every second well had added restriction enzymes.If a band of approximately 160 bp could be seen on the gel, it was concluded as being successful and sent to sequencing.</p>
-<p><img src="https://static.igem.org/mediawiki/2016/6/63/T--SDU-Denmark--Notebook--3_and_.png" alt="" width="461" height="363" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/6/63/T--SDU-Denmark--Notebook--3_and_.png" width="70%" style="margin:auto;"></p>
@@ Line 1,346: / Line 1,346: @@
 <!-------------------------------------------Week 30 ------------------------------------------------------>
 <h4><b> Week 30</b> </h4>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Verification of K2018010 in standard iGEM plasmid (pSB1C3). Date: 16.07.25</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Verification of K2018010 in standard iGEM plasmid (pSB1C3). Date: 16.07.25</button>
 <div id="foo" class="panel">
 <p>The BioBrick containing the gene <a href="http://parts.igem.org/Part:BBa_ K2018010" target="_blank"> K2018010</a> was digested with PstI overnight and with EcoRI the day after by using <a href=" https://static.igem.org/mediawiki/2016/b/b8/T--SDU-Denmark--SOP0017-FastDigest.pdf" target="_blank"> SOP0017 </a>. </p>
@@ Line 1,353: / Line 1,353: @@
 The Gel Photo below shows BG51 (At the photo named; 51). The composite part is 395 bp long. It is possible from the Gel photo to assume that the DNA is cut correctly. From this gel, is the BG51 band of approximately 400 bp purified and thereafter ligated into the standard iGEM plasmid. </p>
-<p><img src="https://static.igem.org/mediawiki/2016/c/cd/T--SDU-Denmark--Notebook--5_and_.png" alt="" width="263" height="275" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/c/cd/T--SDU-Denmark--Notebook--5_and_.png" width="70%" style="margin:auto;"></p>
 <p> After digesting, the sample was ligated by following  <a href="https://static.igem.org/mediawiki/2016/c/c6/T--SDU-Denmark--SOP0015-Ligation.pdf" target="_blank">SOP0015 </a>, with the standard iGEM plasmid, pSB1C3, overnight, and transformed into <em>E.coli Top10</em>.</p>
@@ Line 1,409: / Line 1,409: @@
 <p>The Gel Photo below shows to the left the ladder (green). The following bands are BB54, 1, 2 and 4 which are the ligation of the vector PR17 and the insert BG51. From the photo, it can be seen that sample 54.1 and 54.3 are the vector with the insert, and 54.2 is a religation. </p>
-<p><img src="https://static.igem.org/mediawiki/2016/f/fb/T--SDU-Denmark--Notebook--6_and_.png" alt="" width="263" height="295" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/f/fb/T--SDU-Denmark--Notebook--6_and_.png" width="70%" style="margin:auto;"></p>
 <p>What we had done with the BioBrick<a href="http://parts.igem.org/Part:BBa_K2018010" target="_blank">K2018010</a> can be read in the protocols: <a href="https://static.igem.org/mediawiki/2016/9/9c/T--SDU-Denmark--K2018010-CloningcompositepartintoiGEMstandardplasmidpSB1C3.pdf" target="_blank"> Cloning composite part K2018010 into pSB1C3.</a> and <a href="https://static.igem.org/mediawiki/2016/a/a6/T--SDU-Denmark--IMPACTPurificationofK2018010.pdf" target="_blank">IMPACT Purification of K2018010</a></p>
@@ Line 1,416: / Line 1,416: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Verification of K2018014 in standard iGEM plasmid (pSB1C3). Date: 16.07.25</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Verification of K2018014 in standard iGEM plasmid (pSB1C3). Date: 16.07.25</button>
 <div id="foo" class="panel">
 <p>The BioBrick containing the gene <a href="http://parts.igem.org/Part:BBa_ K2018014" target="_blank"> K2018014</a> was digested with PstI overnight and with EcoRI the day after by using <a href=" https://static.igem.org/mediawiki/2016/b/b8/T--SDU-Denmark--SOP0017-FastDigest.pdf" target="_blank"> SOP0017 </a>. </p>
@@ Line 1,423: / Line 1,423: @@
 The Gel Photo below shows BG52 (At the photo named; 52).The composite part is 458 bp long. It is possible, from the Gel photo, to assume that the DNA is cut correctly. From this gel, the BG52 band of approximately 400 bp purified and thereafter ligated into the standard iGEM plasmid. </p>
-<p><img src="https://static.igem.org/mediawiki/2016/c/cd/T--SDU-Denmark--Notebook--5_and_.png" alt="" width="263" height="275" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/c/cd/T--SDU-Denmark--Notebook--5_and_.png" width="70%" style="margin:auto;"></p>
 <p> After digesting, the sample was ligated by following  <a href="https://static.igem.org/mediawiki/2016/c/c6/T--SDU-Denmark--SOP0015-Ligation.pdf" target="_blank">SOP0015 </a>, with the standard iGEM plasmid, pSB1C3, overnight, and transformed into <em>E.coli Top10</em>.</p>
@@ Line 1,521: / Line 1,521: @@
 <p>The Gel Photo below shows to the left, the ladder (green). The following bands are BB55, 1, 2 and 4 are the ligation of the vector PR17 and the insert BG52. From the photo, it can be assumed that all 3 samples in BB55 are the vector with insert. </p>
-<p><img src="https://static.igem.org/mediawiki/2016/f/fb/T--SDU-Denmark--Notebook--6_and_.png" alt="" width="263" height="295" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/f/fb/T--SDU-Denmark--Notebook--6_and_.png" width="70%" style="margin:auto;"></p>
@@ Line 1,531: / Line 1,531: @@
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Verification of K2018012 in standard iGEM plasmid (pSB1C3). Date: 16.07.26</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Verification of K2018012 in standard iGEM plasmid (pSB1C3). Date: 16.07.26</button>
 <div id="foo" class="panel">
 <p>The BioBrick was ordered from IDT in a plasmid pUCIDT-AMP. When received, it was resuspended to a final concentration at 0.1 µg/µl. </p>
@@ Line 1,542: / Line 1,542: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Verification of K2018015 in standard iGEM plasmid (pSB1C3). Date: 16.07.26</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Verification of K2018015 in standard iGEM plasmid (pSB1C3). Date: 16.07.26</button>
 <div id="foo" class="panel">
 <p>The BioBrick was ordered from IDT in a plasmid pUCIDT-AMP. When received, it was resuspended to a final concentration at 0.1 µg/µl. </p>
@@ Line 1,552: / Line 1,552: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Changes the promoter region on K2018015, and transformation into Top10 <em>E. coli.</em> Date: 16.07.29</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Changes the promoter region on K2018015, and transformation into Top10 <em>E. coli.</em> Date: 16.07.29</button>
 <div id="foo" class="panel">
 <p>The BioBrick, that was ordered from IDT had a defected promoter, and therefore we replaced the promoter region with a functional one. This was done by a phusion PCR, <a href="https://static.igem.org/mediawiki/2016/3/37/T--SDU-Denmark--SOP0010-PhusionPCR.pdf ”target="_blank"> SOP0010 </a>. The primer was; k2018015_IDT. </p>
@@ Line 1,636: / Line 1,636: @@
 <p>Note: There is loaded 5 µl of each sample</p>
-<p><img src="https://static.igem.org/mediawiki/2016/d/d7/T--SDU-Denmark--Notebook--9_and_.png" alt="" width="367" height="231" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/d/d7/T--SDU-Denmark--Notebook--9_and_.png" width="70%" style="margin:auto;"></p>
 <p>What we had done with the BioBrick<a href="http://parts.igem.org/Part:BBa_ K2018015" target="_blank"> K2018015</a> can be read in the protocols: <<a href="https://static.igem.org/mediawiki/2016/c/cc/T--SDU-Denmark--K2018015-CloningcompositepartintoiGEMstandardplasmid.pdf" target="_blank">Cloning composite part K2018015 into pSB1C3</a>and <a href="https://static.igem.org/mediawiki/2016/6/66/T--SDU-Denmark--IMPACTPurificationofK2018015.pdf" target="_blank">IMPACT Purification of K2018015</a> </p>
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Verification of K2018019 in standard iGEM plasmid (pSB1C3). Date: 16.07.25</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Verification of K2018019 in standard iGEM plasmid (pSB1C3). Date: 16.07.25</button>
 <div id="foo" class="panel">
 <p>The BioBrick containing the gene <a href="http://parts.igem.org/Part:BBa_ K2018019" target="_blank"> K2018019</a> was digested with PstI for 2 hours and EcoRI for 30 min. by using <a href=" https://static.igem.org/mediawiki/2016/b/b8/T--SDU-Denmark--SOP0017-FastDigest.pdf" target="_blank"> SOP0017 </a>. </p>
@@ Line 1,648: / Line 1,648: @@
 The two Gel Photos below show the results for BG59 (At the photo named; 59). The composite part is 1739 bp long. It is possible from the Gel photo to assume that the DNA is cut correctly. From this gel, is the BG59 band of around 2000 bp purified and thereafter ligated into the standard iGEM plasmid. </p>
-<p><img src="https://static.igem.org/mediawiki/2016/9/9f/T--SDU-Denmark--Notebook--7_and_.png" alt="" width="542" height="235" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/9/9f/T--SDU-Denmark--Notebook--7_and_.png" width="70%" style="margin:auto;"></p>
 <p> After digesting, the sample was ligated by following  <a href="https://static.igem.org/mediawiki/2016/c/c6/T--SDU-Denmark--SOP0015-Ligation.pdf" target="_blank">SOP0015 </a>, with the standard iGEM plasmid, pSB1C3, overnight, and thereafter transformed, SOP009_v01,  into <em>E.coli Top10</em>.</p>
@@ Line 1,746: / Line 1,746: @@
 <p> Hereafter a colony PCR was runned and verified on a gel. The result is given in the following section: The Gel Photo below shows to the left the ladder (green). The following bands are BB59, 1 to 14, which represent the ligation of the vector BR60 with the insert BG59. From the photo it can be assumed that all the samples are perfectly ligated. One of the colonies can therefore, be used to make ON for the plasmid purification and Freeze stock.</p>
-<p><img src="https://static.igem.org/mediawiki/2016/4/4a/T--SDU-Denmark--Notebook--8_and_.png" alt="" width="432" height="198" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/4/4a/T--SDU-Denmark--Notebook--8_and_.png" width="70%" style="margin:auto;"></p>
 <p>What we had done with the BioBrick<a href="http://parts.igem.org/Part:BBa_K2018019" target="_blank">K2018019</a> can be read in the protocol: <a href="https://static.igem.org/mediawiki/2016/5/5a/T--SDU-Denmark--K2018019-CloningcompositepartintoiGEMstandardplasmid.pdf" target="_blank">Cloning composite part K2018019 into pSB1C3</a></p>
@@ Line 1,755: / Line 1,755: @@
 <h4><b>Week 31</b></h4>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Verification of K2018012 in standard iGEM plasmid (pSB1C3). Date: 16.08.02</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Verification of K2018012 in standard iGEM plasmid (pSB1C3). Date: 16.08.02</button>
 <div id="foo" class="panel">
 <p>The BioBrick containing the gene <a href="http://parts.igem.org/Part:BBa_ K2018012" target="_blank"> K2018012</a> was digested with PstI and XbaI overnight by using <a href=" https://static.igem.org/mediawiki/2016/b/b8/T--SDU-Denmark--SOP0017-FastDigest.pdf" target="_blank"> SOP0017 </a>. </p>
@@ Line 1,856: / Line 1,856: @@
 <p>The following gel picture shows that we successfully cloned our part <a href="http://parts.igem.org/Part:BBa_ K2018012" target="_blank"> K2018012</a>  into the standard iGEM plasmid, pSB1C3. </p>
-<p><img src="https://static.igem.org/mediawiki/2016/c/c6/T--SDU-Denmark--Notebook--10_and_.png" alt="" width="367" height="231" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/c/c6/T--SDU-Denmark--Notebook--10_and_.png" width="70%" style="margin:auto;"></p>
 <p>What we had done with the BioBrick <a href="http://parts.igem.org/Part:BBa_ K2018012" target="_blank"> K2018012</a> can be read in the protocols: <a href="https://static.igem.org/mediawiki/2016/0/09/T--SDU-Denmark--K2018012-CloningcompositepartintoiGEMstandardplasmid.pdf" target="_blank">Cloning composite part K2018012 into pSB1C3</a> and <a href="https://static.igem.org/mediawiki/2016/a/a0/T--SDU-Denmark--IMPACTPurificationofK2018012.pdf" target="_blank">IMPACT Purification of K2018012</a> </p>
@@ Line 1,862: / Line 1,862: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Verification of K2018015 in standard iGEM plasmid (pSB1C3). Date: 16.08.02</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Verification of K2018015 in standard iGEM plasmid (pSB1C3). Date: 16.08.02</button>
 <div id="foo" class="panel">
 <p>The BioBrick containing the gene <a href="http://parts.igem.org/Part:BBa_ K2018015" target="_blank"> K2018015</a> was digested with PstI and XbaI overnight by using <a href=" https://static.igem.org/mediawiki/2016/b/b8/T--SDU-Denmark--SOP0017-FastDigest.pdf" target="_blank"> SOP0017 </a>. </p>
@@ Line 1,963: / Line 1,963: @@
 <p>The Gel Photos below show the first Gel , to the left the ladder (green) can be seen. The following bands are BB79, 1 to 14. On the other gel can be seen first  the ladder (green), and then BB79 sample 15 to 23. BB79 is the ligation of the vector BG17 and the insert BG78. From the photo it can be assumed that all the samples, except; 2 and 6, are perfectly ligated. One of the colonies are therefore used to make ON for plasmid purification and Freeze stock. </p>
-<p><img src="https://static.igem.org/mediawiki/2016/2/21/T--SDU-Denmark--Notebook--11_and_.png" alt="" width="337" height="399" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/2/21/T--SDU-Denmark--Notebook--11_and_.png" width="70%" style="margin:auto;"></p>
 <p>What we had done with the BioBrick<a href="http://parts.igem.org/Part:BBa_ K2018015" target="_blank"> K2018015</a> can be read in the protocols: <<a href="https://static.igem.org/mediawiki/2016/c/cc/T--SDU-Denmark--K2018015-CloningcompositepartintoiGEMstandardplasmid.pdf" target="_blank">Cloning composite part K2018015 into pSB1C3</a>and <a href="https://static.igem.org/mediawiki/2016/6/66/T--SDU-Denmark--IMPACTPurificationofK2018015.pdf" target="_blank">IMPACT Purification of K2018015</a> </p>
@@ Line 1,973: / Line 1,973: @@
   <!-------------------------------------------Week 33 ------------------------------------------------------>
 <h4><b>Week 33</b></h4>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Correct Overhang, D-forward and A-reverse on K2018011. Date: 16.08.19</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Correct Overhang, D-forward and A-reverse on K2018011. Date: 16.08.19</button>
 <div id="foo" class="panel">
 <p><a href="http://parts.igem.org/Part:BBa_ K2018011" target="_blank"> K2018011</a>in standard iGEM plasmid (pSB1C3) was used in a phusion PCR, <a href="https://static.igem.org/mediawiki/2016/3/37/T--SDU-Denmark--SOP0010-PhusionPCR.pdf ”target="_blank"> SOP0010 </a>. The primes have an overhang, which makes the gene compatible with the silk overhang, and thereby enabling us to make a hybrid silk fiber.</p>
@@ Line 2,028: / Line 2,028: @@
 <p> The result is verified by Gel electrophoresis. The result is shown here:</p>
-<p><img src="https://static.igem.org/mediawiki/2016/6/63/T--SDU-Denmark--Notebook--12_and_.png" alt="" width="452" height="288" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/6/63/T--SDU-Denmark--Notebook--12_and_.png" width="70%" style="margin:auto;"></p>
 <p>Lane no. 5 is ThuricinS with DA-overhangs and is expected to be 217 bp long. We have used the yellow ladder, where the fourth band represents 200 bp. Based on that, it can be assumed that we succeeded at getting silk-overhangs on ThuricinS.</p>
@@ Line 2,037: / Line 2,037: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Correct Overhang, D-forward and E-reverse on K2018011. Date: 16.08.19</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Correct Overhang, D-forward and E-reverse on K2018011. Date: 16.08.19</button>
 <div id="foo" class="panel">
 <p><a href="http://parts.igem.org/Part:BBa_ K2018011" target="_blank"> K2018011</a>in standard iGEM plasmid (pSB1C3) was used in a phusion PCR, <a href="https://static.igem.org/mediawiki/2016/3/37/T--SDU-Denmark--SOP0010-PhusionPCR.pdf ”target="_blank"> SOP0010 </a>. The primes have an overhang, which makes the gene compatible with the silk overhang, and thereby enabling us to make a hybrid silk fiber.</p>
@@ Line 2,092: / Line 2,092: @@
 <p> The result is verified by Gel electrophoresis. The result is shown here:</p>
-<p><img src="https://static.igem.org/mediawiki/2016/a/a8/T--SDU-Denmark--Notebook--13_and_.png" alt="" width="468" height="327" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/a/a8/T--SDU-Denmark--Notebook--13_and_.png" width="70%" style="margin:auto;"></p>
 <p>The lane marked no. 2 is ThuricinS with DE-overhangs and it is assumed to be 217 bp long. We have used the yellow ladder, where the fourth band is 200 bp long. Based on that, it can be assumed that we succeeded at getting silk-overhangs on ThuricinS.</p>
@@ Line 2,101: / Line 2,101: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Correct Overhang, E-forward and A-reverse on K2018011. Date: 16.08.19</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Correct Overhang, E-forward and A-reverse on K2018011. Date: 16.08.19</button>
 <div id="foo" class="panel">
 <p><a href="http://parts.igem.org/Part:BBa_ K2018011" target="_blank"> K2018011</a>in standard iGEM plasmid (pSB1C3) was used in a phusion PCR, <a href="https://static.igem.org/mediawiki/2016/3/37/T--SDU-Denmark--SOP0010-PhusionPCR.pdf ”target="_blank"> SOP0010 </a>. The primes have an overhang, which makes the gene compatible with the silk overhang, and thereby enabling us to make a hybrid silk fiber.</p>
@@ Line 2,156: / Line 2,156: @@
 <p> The result is verified by Gel electrophoresis. The result is shown here:</p>
-<p><img src="https://static.igem.org/mediawiki/2016/6/63/T--SDU-Denmark--Notebook--12_and_.png" alt="" width="452" height="288" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/6/63/T--SDU-Denmark--Notebook--12_and_.png" width="70%" style="margin:auto;"></p>
 <p>The lane marked no. 7 and 8 are ThuricinS with EA-overhangs and it are expected to be 217 bp long. We have used the yellow ladder, where the fourth band is 200 bp long. Based on that, it can be expected that we succeeded at getting silk-overhangs on ThuricinS.</p>
@@ Line 2,165: / Line 2,165: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Correct Overhang, E-forward and D-reverse on K2018011. Date: 16.08.19</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Correct Overhang, E-forward and D-reverse on K2018011. Date: 16.08.19</button>
 <div id="foo" class="panel">
 <p><a href="http://parts.igem.org/Part:BBa_ K2018011" target="_blank"> K2018011</a>in standard iGEM plasmid (pSB1C3) was used in a phusion PCR, <a href="https://static.igem.org/mediawiki/2016/3/37/T--SDU-Denmark--SOP0010-PhusionPCR.pdf ”target="_blank"> SOP0010 </a>. The primes have an overhang, which makes the gene compatible with the silk overhang, and thereby enabling us to make a hybrid silk fiber.</p>
@@ Line 2,220: / Line 2,220: @@
 <p> The result is verified by Gel electrophoresis. The result is shown here:</p>
-<p><img src="https://static.igem.org/mediawiki/2016/6/63/T--SDU-Denmark--Notebook--12_and_.png" alt="" width="452" height="288" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/6/63/T--SDU-Denmark--Notebook--12_and_.png" width="70%" style="margin:auto;"></p>
 <p>The lane marked no. 4 and 8 are ThuricinS with ED-overhangs and is expected to be 217 bp long. We have used the yellow ladder, where the fourth band is 200 bp. Based on that it  can be assumed that we succeeded at getting silk-overhangs on ThuricinS.</p>
@@ Line 2,231: / Line 2,231: @@
 <h4><b>Week 34</b></h4>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Correct Overhang, to IMPACT on K2018010. Date: 16.08.25</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Correct Overhang, to IMPACT on K2018010. Date: 16.08.25</button>
 <div id="foo" class="panel">
 <p>For using the IMPACT method for purification of bacteriocins, some specific restrictions sites are necessary. Therefore a Phusion PCR was made by following <a href="https://static.igem.org/mediawiki/2016/3/37/T--SDU-Denmark--SOP0010-PhusionPCR.pdf ”target="_blank"> SOP0010 </a>. </p>
@@ Line 2,289: / Line 2,289: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Correct Overhang, to IMPACT on K2018011. Date: 16.08.25</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Correct Overhang, to IMPACT on K2018011. Date: 16.08.25</button>
 <div id="foo" class="panel">
 <p>For using the IMPACT method for purification of bacteriocins, some specific restrictions sites are necessary. Therefore a Phusion PCR was made by following <a href="https://static.igem.org/mediawiki/2016/3/37/T--SDU-Denmark--SOP0010-PhusionPCR.pdf ”target="_blank"> SOP0010 </a>. </p>
@@ Line 2,349: / Line 2,349: @@
 <!-------------------------------------------Week 35------------------------------------------------------>
 <h4><b>Week 35</b></h4>
-  <button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Verification of K2018011 with D-forward and A-reverse overhang, in standard iGEM plasmid (pSB1C3). Date: 16.09.01</button>
+  <button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Verification of K2018011 with D-forward and A-reverse overhang, in standard iGEM plasmid (pSB1C3). Date: 16.09.01</button>
 <div id="foo" class="panel">
 <p>The BioBrick was digested with with SpeI for 2h  and EcoRI for 30 min, by following <a href=" https://static.igem.org/mediawiki/2016/b/b8/T--SDU-Denmark--SOP0017-FastDigest.pdf" target="_blank"> SOP0017 </a>. </p>
@@ Line 2,449: / Line 2,449: @@
 <p>ThuricinS with silk overhangs, together with primer overhangs (VF2 and VR), is 517 bp long. Fermentas Generuler 1 kb ladder plus was used, so from these pictures it can verified that ThuricinS with DA overhangs have the right length. </p>
-<p><img src="https://static.igem.org/mediawiki/2016/1/12/T--SDU-Denmark--Notebook--14_and_.png" alt="" width="491" height="255" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/1/12/T--SDU-Denmark--Notebook--14_and_.png" width="70%" style="margin:auto;"></p>
 <p>What we had done with the BioBrick <a href="http://parts.igem.org/Part:BBa_K2018011" target="_blank">K2018011</a> can be read in the protocols: <a href="https://static.igem.org/mediawiki/2016/e/ea/T--SDU-Denmark--K2018011clonedintopSB1C3.pdf" target="_blank">Cloning composite part K2018011 into pSB1C3</a>,  <a href="https://static.igem.org/mediawiki/2016/1/12/T--SDU-Denmark--CloningThuricinwithDAsilk-overhangsintopSB1C3.pdf" target="_blank">Cloning ThuricinS with DA silk-overhangs into pSB1C3</a>, <a href="https://static.igem.org/mediawiki/2016/0/0d/T--SDU-Denmark--CloningThuricinwithDEsilk-overhangsintopSB1C3.pdf" target="_blank">Cloning ThuricinS with DE silk-overhangs into pSB1C3</a>, <a href="https://static.igem.org/mediawiki/2016/5/52/T--SDU-Denmark--CloningThuricinwithEAsilk-overhangsintopSB1C3.pdf" target="_blank">Cloning ThuricinS with EA silk-overhangs into pSB1C3</a>, <a href="https://static.igem.org/mediawiki/2016/b/b1/T--SDU-Denmark--CloningThuricinwithEDsilk-overhangsintopSB1C3.pdf" target="_blank">Cloning ThuricinS with ED silk-overhangs into pSB1C3</a> and <a href="https://static.igem.org/mediawiki/2016/0/03/T--SDU-Denmark--IMPACTPurificationofK2018011.pdf" target="_blank">IMPACT Purification of K2018011</a> </p>
@@ Line 2,456: / Line 2,456: @@
 </div>
-  <button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Verification of K2018011 with D-forward and E-reverse overhang, in standard iGEM plasmid (pSB1C3). Date: 16.09.01</button>
+  <button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Verification of K2018011 with D-forward and E-reverse overhang, in standard iGEM plasmid (pSB1C3). Date: 16.09.01</button>
 <div id="foo" class="panel">
 <p>The BioBrick was digested with with SpeI for 2h  and EcoRI for 30 min, by following <a href=" https://static.igem.org/mediawiki/2016/b/b8/T--SDU-Denmark--SOP0017-FastDigest.pdf" target="_blank"> SOP0017 </a>. </p>
@@ Line 2,556: / Line 2,556: @@
 <p>ThuricinS with silk overhangs, together with primer overhangs (VF2 and VR), is 517 bp long. Fermentas Generuler 1 kb ladder plus was used, so from these pictures it can verified that ThuricinS with DE overhangs have the right length. </p>
-<p><img src="https://static.igem.org/mediawiki/2016/c/cd/T--SDU-Denmark--Notebook--15_and_.png" alt="" width="224" height="233" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/c/cd/T--SDU-Denmark--Notebook--15_and_.png" width="70%" style="margin:auto;"></p>
 <p>What we had done with the BioBrick <a href="http://parts.igem.org/Part:BBa_K2018011" target="_blank">K2018011</a> can be read in the protocols: <a href="https://static.igem.org/mediawiki/2016/e/ea/T--SDU-Denmark--K2018011clonedintopSB1C3.pdf" target="_blank">Cloning composite part K2018011 into pSB1C3</a>,  <a href="https://static.igem.org/mediawiki/2016/1/12/T--SDU-Denmark--CloningThuricinwithDAsilk-overhangsintopSB1C3.pdf" target="_blank">Cloning ThuricinS with DA silk-overhangs into pSB1C3</a>, <a href="https://static.igem.org/mediawiki/2016/0/0d/T--SDU-Denmark--CloningThuricinwithDEsilk-overhangsintopSB1C3.pdf" target="_blank">Cloning ThuricinS with DE silk-overhangs into pSB1C3</a>, <a href="https://static.igem.org/mediawiki/2016/5/52/T--SDU-Denmark--CloningThuricinwithEAsilk-overhangsintopSB1C3.pdf" target="_blank">Cloning ThuricinS with EA silk-overhangs into pSB1C3</a>, <a href="https://static.igem.org/mediawiki/2016/b/b1/T--SDU-Denmark--CloningThuricinwithEDsilk-overhangsintopSB1C3.pdf" target="_blank">Cloning ThuricinS with ED silk-overhangs into pSB1C3</a> and <a href="https://static.igem.org/mediawiki/2016/0/03/T--SDU-Denmark--IMPACTPurificationofK2018011.pdf" target="_blank">IMPACT Purification of K2018011</a> </p>
@@ Line 2,563: / Line 2,563: @@
 </div>
-  <button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Verification of K2018011 with E-forward and A-reverse overhang, in standard iGEM plasmid (pSB1C3). Date: 16.09.01</button>
+  <button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Verification of K2018011 with E-forward and A-reverse overhang, in standard iGEM plasmid (pSB1C3). Date: 16.09.01</button>
 <div id="foo" class="panel">
 <p>The BioBrick was digested with with SpeI for 2h  and EcoRI for 30 min, by following <a href=" https://static.igem.org/mediawiki/2016/b/b8/T--SDU-Denmark--SOP0017-FastDigest.pdf" target="_blank"> SOP0017 </a>. </p>
@@ Line 2,663: / Line 2,663: @@
 <p>ThuricinS with silk overhangs, together with primer overhangs (VF2 and VR), is 517 bp long. Fermentas Generuler 1 kb ladder plus was used, so from these pictures it can verified that ThuricinS with EA overhangs have the right length. </p>
-<p><img src="https://static.igem.org/mediawiki/2016/c/cd/T--SDU-Denmark--Notebook--15_and_.png" alt="" width="224" height="233" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/c/cd/T--SDU-Denmark--Notebook--15_and_.png" width="70%" style="margin:auto;"/></p>
 <p>What we had done with the BioBrick <a href="http://parts.igem.org/Part:BBa_K2018011" target="_blank">K2018011</a> can be read in the protocols: <a href="https://static.igem.org/mediawiki/2016/e/ea/T--SDU-Denmark--K2018011clonedintopSB1C3.pdf" target="_blank">Cloning composite part K2018011 into pSB1C3</a>,  <a href="https://static.igem.org/mediawiki/2016/1/12/T--SDU-Denmark--CloningThuricinwithDAsilk-overhangsintopSB1C3.pdf" target="_blank">Cloning ThuricinS with DA silk-overhangs into pSB1C3</a>, <a href="https://static.igem.org/mediawiki/2016/0/0d/T--SDU-Denmark--CloningThuricinwithDEsilk-overhangsintopSB1C3.pdf" target="_blank">Cloning ThuricinS with DE silk-overhangs into pSB1C3</a>, <a href="https://static.igem.org/mediawiki/2016/5/52/T--SDU-Denmark--CloningThuricinwithEAsilk-overhangsintopSB1C3.pdf" target="_blank">Cloning ThuricinS with EA silk-overhangs into pSB1C3</a>, <a href="https://static.igem.org/mediawiki/2016/b/b1/T--SDU-Denmark--CloningThuricinwithEDsilk-overhangsintopSB1C3.pdf" target="_blank">Cloning ThuricinS with ED silk-overhangs into pSB1C3</a> and <a href="https://static.igem.org/mediawiki/2016/0/03/T--SDU-Denmark--IMPACTPurificationofK2018011.pdf" target="_blank">IMPACT Purification of K2018011</a> </p>
@@ Line 2,670: / Line 2,670: @@
 </div>
-  <button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Verification of K2018011 with E-forward and D-reverse overhang, in standard iGEM plasmid (pSB1C3). Date: 16.09.01</button>
+  <button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Verification of K2018011 with E-forward and D-reverse overhang, in standard iGEM plasmid (pSB1C3). Date: 16.09.01</button>
 <div id="foo" class="panel">
 <p>The BioBrick was digested with with SpeI for 2h  and EcoRI for 30 min, by following <a href=" https://static.igem.org/mediawiki/2016/b/b8/T--SDU-Denmark--SOP0017-FastDigest.pdf" target="_blank"> SOP0017 </a>. </p>
@@ Line 2,770: / Line 2,770: @@
 <p>ThuricinS with silk overhangs, together with primer overhangs (VF2 and VR), is 517 bp long. Fermentas Generuler 1 kb ladder plus was used, so from these pictures it can verified that ThuricinS with ED overhangs have the right length. </p>
-<p><img src="https://static.igem.org/mediawiki/2016/6/6a/T--SDU-Denmark--Notebook--16_and_.png" alt="" width="410" height="365" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/6/6a/T--SDU-Denmark--Notebook--16_and_.png" width="70%" style="margin:auto;"></p>
 <p>What we had done with the BioBrick <a href="http://parts.igem.org/Part:BBa_K2018011" target="_blank">K2018011</a> can be read in the protocols: <a href="https://static.igem.org/mediawiki/2016/e/ea/T--SDU-Denmark--K2018011clonedintopSB1C3.pdf" target="_blank">Cloning composite part K2018011 into pSB1C3</a>,  <a href="https://static.igem.org/mediawiki/2016/1/12/T--SDU-Denmark--CloningThuricinwithDAsilk-overhangsintopSB1C3.pdf" target="_blank">Cloning ThuricinS with DA silk-overhangs into pSB1C3</a>, <a href="https://static.igem.org/mediawiki/2016/0/0d/T--SDU-Denmark--CloningThuricinwithDEsilk-overhangsintopSB1C3.pdf" target="_blank">Cloning ThuricinS with DE silk-overhangs into pSB1C3</a>, <a href="https://static.igem.org/mediawiki/2016/5/52/T--SDU-Denmark--CloningThuricinwithEAsilk-overhangsintopSB1C3.pdf" target="_blank">Cloning ThuricinS with EA silk-overhangs into pSB1C3</a>, <a href="https://static.igem.org/mediawiki/2016/b/b1/T--SDU-Denmark--CloningThuricinwithEDsilk-overhangsintopSB1C3.pdf" target="_blank">Cloning ThuricinS with ED silk-overhangs into pSB1C3</a> and <a href="https://static.igem.org/mediawiki/2016/0/03/T--SDU-Denmark--IMPACTPurificationofK2018011.pdf" target="_blank">IMPACT Purification of K2018011</a> </p>
@@ Line 2,778: / Line 2,778: @@
 <!-------------------------------------------Week 36 ------------------------------------------------------>
 <h4><b>Week 36</b></h4>
-  <button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" height="20%" style="float:right;margin-left:15px;">Verification of K2018043, in standard iGEM plasmid (pSB1C3). Date: 16.09.10</button>
+  <button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" width="70%" style="margin:auto;">Verification of K2018043, in standard iGEM plasmid (pSB1C3). Date: 16.09.10</button>
 <div id="foo" class="panel">
 <p>The BioBrick was first digested with restrictions enzymes by following <a href=" https://static.igem.org/mediawiki/2016/b/b8/T--SDU-Denmark--SOP0017-FastDigest.pdf" target="_blank"> SOP0017 </a>. The restriction enzymes were: PstI and XbaI for 2h. After digesting, the sample was verified by Gel electrophoresis.  </p>
@@ Line 2,919: / Line 2,919: @@
 <p>The Gel Photo below shows the cPCR results. VR and VF2 primers adds a total of 300 bp, the gene is 119 bp, which gives a total DNA length of 419 bp.  From this gel the ligation  of the gen incorporated into psB1C3 can be observed.</p>
-<p><img src="https://static.igem.org/mediawiki/2016/c/c8/T--SDU-Denmark--Notebook--17_and_.png" alt="" width="500" height="287" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/c/c8/T--SDU-Denmark--Notebook--17_and_.png" width="70%" style="margin:auto;"></p>
 <p>This is the picture of the colony PCR where 1.2 to 1.6 are different colonies of the same plate for MaSp1 CD. 1.3 was used to purify plasmids. </p>
 <p>The second gel photo shows cutting of SB100 with Xbal and PstI. Every second well had added restriction enzymes. If a band of 145 bp long was observed on the gel of ligation and transformation of SB100, it was concluded successful and sent to sequencing. </p>
-<p><img src="https://static.igem.org/mediawiki/2016/5/57/T--SDU-Denmark--Notebook--18_and_.png" alt="" width="508" height="319" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/5/57/T--SDU-Denmark--Notebook--18_and_.png" width="70%" style="margin:auto;"></p>
 <p>What we have done with the BioBrick <a href="http://parts.igem.org/Part:BBa_K2018043" target="_blank">K2018043</a> can be read in the protocol: ><a href="https://static.igem.org/mediawiki/2016/3/3f/T--SDU-Denmark--K2018043-CloningBasicpartintoiGEMstandardplasmidpSB1C3.pdf" target="_blank">Cloning Basic part K2018043 into pSB1C3</a> </p>
@@ Line 2,932: / Line 2,932: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" height="20%" style="float:right;margin-left:15px;">Verification of K2018044, in standard iGEM plasmid (pSB1C3). Date: 16.09.10</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/4/45/T--SDU-Denmark--minisilk.png" width="70%" style="margin:auto;">Verification of K2018044, in standard iGEM plasmid (pSB1C3). Date: 16.09.10</button>
 <div id="foo" class="panel">
 <p>The BioBrick was first digested with restrictions enzymes by following <a href=" https://static.igem.org/mediawiki/2016/b/b8/T--SDU-Denmark--SOP0017-FastDigest.pdf" target="_blank"> SOP0017 </a>. The restriction enzymes were: PstI and XbaI for 2 h. After digesting, the sample was verified by Gel electrophoresis. </p>
@@ Line 3,073: / Line 3,073: @@
 <p>The Gel Photo below shows the cPCR results. VR and VF2 primers adds a total of 300 bp, the gene is 119 bp long, which gives a total DNA length of 419 bp.  From this gel the ligation  of the gen incorporated into psB1C3 can be indicated .</p>
-<p><img src="https://static.igem.org/mediawiki/2016/c/c8/T--SDU-Denmark--Notebook--17_and_.png" alt="" width="500" height="287" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/c/c8/T--SDU-Denmark--Notebook--17_and_.png" width="70%" style="margin:auto;"></p>
 <p>This is the picture of the colony PCR where 2.1 to 2.3 are different colonies of the same plate for MaSp2 CD. 2.1 was used to purify plasmids </p>
 <p>The second gel photo shows cutting of SB101 with Xbal and PstI. Every second well had added restriction enzymes. From this gel, the ligation and transformation of SB101, a band around 145 bp after digestion, was concluded as being successful and sent to sequencing. </p>
-<p><img src="https://static.igem.org/mediawiki/2016/5/57/T--SDU-Denmark--Notebook--18_and_.png" alt="" width="508" height="319" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/5/57/T--SDU-Denmark--Notebook--18_and_.png" width="70%" style="margin:auto;"></p>
 <p>What we have done with the BioBrick <a href="http://parts.igem.org/Part:BBa_K2018044" target="_blank">K2018044</a> can be read in the protocol:  <a href="https://static.igem.org/mediawiki/2016/1/19/T--SDU-Denmark--K2018044-CloningBasicpartintoiGEMstandardplasmidpsB1C3.pdf" target="_blank">Cloning Basic part K2018044 into pSB1C3</a> </p>
@@ Line 3,085: / Line 3,085: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Successful transformation to the IMPACT vector.  K2018010. Date: 16.09.08</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Successful transformation to the IMPACT vector.  K2018010. Date: 16.09.08</button>
 <div id="foo" class="panel">
 <p>Before purifying the bacteriocin by IMPACT, the bacteriocin must first be transformed into the IMPACT vector bacteria,  <em>E. Coli:ER2566</em>.  To do this, we  incorporated  the  pPCR product into the IMPACT plasmid, pTXB1. Thereafter  the products are digested by following the <a href="https://static.igem.org/mediawiki/2016/d/d9/T--SDU-Denmark--iGEM2016SOP0025_v01-Impact_Purification.pdf" target="_blank">SOP0025.</a> step. 8.1.1. and afterwards ligated by following step 8.1.3 → 8.1.4 in the same SOP. </p>
@@ Line 3,092: / Line 3,092: @@
 <p>The following gel image were used to verify that <a href="http://parts.igem.org/Part:BBa_K2018010 " target="_blank">K2018010 </a>were successfully cloned into pTXB1 (the IMPACT vector). Lane 4-6 is K2018010/pTXB1. Lane 4 and 6 showed the correct sized band. Lane 5 is a negative. </p>
-<p><img src="https://static.igem.org/mediawiki/2016/d/de/T--SDU-Denmark--Notebook--19_and_.png" alt="" width="632" height="208" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/d/de/T--SDU-Denmark--Notebook--19_and_.png" width="70%" style="margin:auto;"></p>
 <p>What we had done with the BioBrick<a href="http://parts.igem.org/Part:BBa_K2018010" target="_blank">K2018010</a> can be read in the protocols: <a href="https://static.igem.org/mediawiki/2016/9/9c/T--SDU-Denmark--K2018010-CloningcompositepartintoiGEMstandardplasmidpSB1C3.pdf" target="_blank"> Cloning composite part K2018010 into pSB1C3.</a> and <a href="https://static.igem.org/mediawiki/2016/a/a6/T--SDU-Denmark--IMPACTPurificationofK2018010.pdf" target="_blank">IMPACT Purification of K2018010</a></p>
@@ Line 3,099: / Line 3,099: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Successful transformation to the IMPACT vector.  K2018011. Date: 16.09.08-</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Successful transformation to the IMPACT vector.  K2018011. Date: 16.09.08-</button>
 <div id="foo" class="panel">
 <p>Before purifying the bacteriocin by IMPACT, the bacteriocin must first be transformed into the IMPACT vector bacteria,  <em>E. Coli:ER2566</em>.  To do this, we  incorporated  the  pPCR product into the IMPACT plasmid, pTXB1. Thereafter  the products are digested by following the <a href="https://static.igem.org/mediawiki/2016/d/d9/T--SDU-Denmark--iGEM2016SOP0025_v01-Impact_Purification.pdf" target="_blank">SOP0025.</a> step. 8.1.1. and afterwards ligated by following step 8.1.3 → 8.1.4 in the same SOP. </p>
@@ Line 3,106: / Line 3,106: @@
 <p>The following gel image were used to verify that <a href="http://parts.igem.org/Part:BBa_K2018011 " target="_blank">K2018011 </a>were successfully cloned into pTXB1 (the IMPACT vector). Lane 4-6 is K2018010/pTXB1. Lane 1-3  is K2018011/pTXB1 and all showed the correct sized band. </p>
-<p><img src="https://static.igem.org/mediawiki/2016/d/de/T--SDU-Denmark--Notebook--19_and_.png" alt="" width="632" height="208" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/d/de/T--SDU-Denmark--Notebook--19_and_.png" width="70%" style="margin:auto;"></p>
 <p>What we had done with the BioBrick <a href="http://parts.igem.org/Part:BBa_K2018011" target="_blank">K2018011</a> can be read in the protocols: <a href="https://static.igem.org/mediawiki/2016/e/ea/T--SDU-Denmark--K2018011clonedintopSB1C3.pdf" target="_blank">Cloning composite part K2018011 into pSB1C3</a>,  <a href="https://static.igem.org/mediawiki/2016/1/12/T--SDU-Denmark--CloningThuricinwithDAsilk-overhangsintopSB1C3.pdf" target="_blank">Cloning ThuricinS with DA silk-overhangs into pSB1C3</a>, <a href="https://static.igem.org/mediawiki/2016/0/0d/T--SDU-Denmark--CloningThuricinwithDEsilk-overhangsintopSB1C3.pdf" target="_blank">Cloning ThuricinS with DE silk-overhangs into pSB1C3</a>, <a href="https://static.igem.org/mediawiki/2016/5/52/T--SDU-Denmark--CloningThuricinwithEAsilk-overhangsintopSB1C3.pdf" target="_blank">Cloning ThuricinS with EA silk-overhangs into pSB1C3</a>, <a href="https://static.igem.org/mediawiki/2016/b/b1/T--SDU-Denmark--CloningThuricinwithEDsilk-overhangsintopSB1C3.pdf" target="_blank">Cloning ThuricinS with ED silk-overhangs into pSB1C3</a> and <a href="https://static.igem.org/mediawiki/2016/0/03/T--SDU-Denmark--IMPACTPurificationofK2018011.pdf" target="_blank">IMPACT Purification of K2018011</a> </p>
@@ Line 3,113: / Line 3,113: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Successful transformation to the IMPACT vector.  K2018012. Date: 16.09.08</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Successful transformation to the IMPACT vector.  K2018012. Date: 16.09.08</button>
 <div id="foo" class="panel">
 <p>Before purifying the bacteriocin by IMPACT, the bacteriocin must first be transformed into the IMPACT vector bacteria,  <em>E. Coli:ER2566</em>.  To do this, we  incorporated  the  pPCR product into the IMPACT plasmid, pTXB1. Thereafter  the products are digested by following the <a href="https://static.igem.org/mediawiki/2016/d/d9/T--SDU-Denmark--iGEM2016SOP0025_v01-Impact_Purification.pdf" target="_blank">SOP0025.</a> step. 8.1.1. and afterwards ligated by following step 8.1.3 → 8.1.4 in the same SOP. </p>
@@ Line 3,119: / Line 3,119: @@
 <p>The following gel image were used to verify that <a href="http://parts.igem.org/Part:BBa_K2018012 " target="_blank">K2018012 </a>were successfully cloned into pTXB1 (the IMPACT vector). Lane 9-12 is K2018012/pTXB1 and they all showed the correct sized band. </p>
-<p><img src="https://static.igem.org/mediawiki/2016/6/6b/T--SDU-Denmark--Notebook--20_and_.png" alt="" width="646" height="211" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/6/6b/T--SDU-Denmark--Notebook--20_and_.png" width="70%" style="margin:auto;"></p>
 <p>What we had done with the BioBrick <a href="http://parts.igem.org/Part:BBa_K2018012" target="_blank">K2018012</a> can be read in the protocols: <a href="https://static.igem.org/mediawiki/2016/0/09/T--SDU-Denmark--K2018012-CloningcompositepartintoiGEMstandardplasmid.pdf" target="_blank">Cloning composite part K2018012 into pSB1C3</a> and <a href="https://static.igem.org/mediawiki/2016/a/a0/T--SDU-Denmark--IMPACTPurificationofK2018012.pdf" target="_blank">IMPACT Purification of K2018012</a> </p>
@@ Line 3,125: / Line 3,125: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Successful transformation to the IMPACT vector.  K2018014. Date: 16.09.08</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Successful transformation to the IMPACT vector.  K2018014. Date: 16.09.08</button>
 <div id="foo" class="panel">
 <p>Before purifying the bacteriocin by IMPACT, the bacteriocin must first be transformed into the IMPACT vector bacteria,  <em>E. Coli:ER2566</em>.  To do this, we  incorporated  the  pPCR product into the IMPACT plasmid, pTXB1. Thereafter  the products are digested by following the <a href="https://static.igem.org/mediawiki/2016/d/d9/T--SDU-Denmark--iGEM2016SOP0025_v01-Impact_Purification.pdf" target="_blank">SOP0025.</a> step. 8.1.1. and afterwards ligated by following step 8.1.3 → 8.1.4 in the same SOP. </p>
@@ Line 3,131: / Line 3,131: @@
 <p>The following gel image were used to verify that <a href="http://parts.igem.org/Part:BBa_K2018014 " target="_blank">K2018014 </a>were successfully cloned into pTXB1 (the IMPACT vector). The three first lanes is K2018014/pTXB1, where lane 1 and 3 showed the correct sized band but lane 2 was negative.. </p>
-<p><img src="https://static.igem.org/mediawiki/2016/d/de/T--SDU-Denmark--Notebook--19_and_.png" alt="" width="632" height="208" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/d/de/T--SDU-Denmark--Notebook--19_and_.png" width="70%" style="margin:auto;"></p>
 <p>What we had done with the BioBrick<a href="http://parts.igem.org/Part:BBa_K2018014" target="_blank">K2018014</a> can be read in the protocols: <a href="https://static.igem.org/mediawiki/2016/b/bf/T--SDU-Denmark--K2018014-CloningcompositepartintoiGEMstandardplasmid.pdf" target="_blank">Cloning composite part K2018014 into pSB1C3</a> and <a href="https://static.igem.org/mediawiki/2016/f/f6/T--SDU-Denmark--IMPACTPurificationofK2018014.pdf" target="_blank">IMPACT Purification of K2018014</a></p>
@@ Line 3,137: / Line 3,137: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%"style="float:right;margin-left:15px;">Successful transformation to the IMPACT vector.  K2018015. Date: 16.09.08</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Successful transformation to the IMPACT vector.  K2018015. Date: 16.09.08</button>
 <div id="foo" class="panel">
 <p>Before purifying the bacteriocin by IMPACT, the bacteriocin must first be transformed into the IMPACT vector bacteria,  <em>E. Coli:ER2566</em>.  To do this, we  incorporated  the  pPCR product into the IMPACT plasmid, pTXB1. Thereafter  the products are digested by following the <a href="https://static.igem.org/mediawiki/2016/d/d9/T--SDU-Denmark--iGEM2016SOP0025_v01-Impact_Purification.pdf" target="_blank">SOP0025.</a> step. 8.1.1. and afterwards ligated by following step 8.1.3 → 8.1.4 in the same SOP. </p>
@@ Line 3,143: / Line 3,143: @@
 <p>The following gel image were used to verify that <a href="http://parts.igem.org/Part:BBa_K2018015 " target="_blank">K2018015 </a>were successfully cloned into pTXB1 (the IMPACT vector). We used three different primers to ‘cross amplify’ the gene and a part of the vector plasmid, getting 2 different DNA bands. All of the given results were, as expected, at 330 bp and 494 bp. </p>
-<p><img src="https://static.igem.org/mediawiki/2016/1/14/T--SDU-Denmark--Notebook--21_and_.png" alt="" width="502" height="125" /></p>
+<p><img src="https://static.igem.org/mediawiki/2016/1/14/T--SDU-Denmark--Notebook--21_and_.png" width="70%" style="margin:auto;"></p>
 <p>What we had done with the BioBrick<a href="http://parts.igem.org/Part:BBa_ K2018015" target="_blank"> K2018015</a> can be read in the protocols: <<a href="https://static.igem.org/mediawiki/2016/c/cc/T--SDU-Denmark--K2018015-CloningcompositepartintoiGEMstandardplasmid.pdf" target="_blank">Cloning composite part K2018015 into pSB1C3</a>and <a href="https://static.igem.org/mediawiki/2016/6/66/T--SDU-Denmark--IMPACTPurificationofK2018015.pdf" target="_blank">IMPACT Purification of K2018015</a> </p>
@@ Line 3,152: / Line 3,152: @@
 <!-------------------------------------------Week 38 ------------------------------------------------------>
 <h4><b>Week 38</b></h4>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Successful purification by IMPACT.  K2018010. Date: 16.09.25</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Successful purification by IMPACT.  K2018010. Date: 16.09.25</button>
 <div id="foo" class="panel">
 <p>The IMPACT purification is done by following the <a href="https://static.igem.org/mediawiki/2016/d/d9/T--SDU-Denmark--iGEM2016SOP0025_v01-Impact_Purification.pdf" target="_blank">SOP0025.</a> step.  8.1.6→ 8.4.3. and the purity is determined by Bradford Protein Concentration determination. </p>
@@ Line 3,163: / Line 3,163: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Successful purification by IMPACT.  K2018011. Date: 16.09.25</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Successful purification by IMPACT.  K2018011. Date: 16.09.25</button>
 <div id="foo" class="panel">
 <p>The IMPACT purification is done by following the <a href="https://static.igem.org/mediawiki/2016/d/d9/T--SDU-Denmark--iGEM2016SOP0025_v01-Impact_Purification.pdf" target="_blank">SOP0025.</a> step.  8.1.6→ 8.4.3. and the purity is determined by Bradford Protein Concentration determination. </p>
@@ Line 3,174: / Line 3,174: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Successful purification by IMPACT.  K2018012. Date: 16.09.25</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Successful purification by IMPACT.  K2018012. Date: 16.09.25</button>
 <div id="foo" class="panel">
 <p>The IMPACT purification is done by following the <a href="https://static.igem.org/mediawiki/2016/d/d9/T--SDU-Denmark--iGEM2016SOP0025_v01-Impact_Purification.pdf" target="_blank">SOP0025.</a> step.  8.1.6→ 8.4.3. and the purity is determined by Bradford Protein Concentration determination. </p>
@@ Line 3,184: / Line 3,184: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Successful purification by IMPACT.  K2018014. Date: 16.09.25</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Successful purification by IMPACT.  K2018014. Date: 16.09.25</button>
 <div id="foo" class="panel">
 <p>The IMPACT purification is done by following the <a href="https://static.igem.org/mediawiki/2016/d/d9/T--SDU-Denmark--iGEM2016SOP0025_v01-Impact_Purification.pdf" target="_blank">SOP0025.</a> step.  8.1.6→ 8.4.3. and the purity is determined by Bradford Protein Concentration determination. </p>
@@ Line 3,194: / Line 3,194: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Successful purification by IMPACT.  K2018015. Date: 16.09.25</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Successful purification by IMPACT.  K2018015. Date: 16.09.25</button>
 <div id="foo" class="panel">
 <p>The IMPACT purification is done by following the <a href="https://static.igem.org/mediawiki/2016/d/d9/T--SDU-Denmark--iGEM2016SOP0025_v01-Impact_Purification.pdf" target="_blank">SOP0025.</a> step.  8.1.6→ 8.4.3. and the purity is determined by Bradford Protein Concentration determination. </p>
@@ Line 3,209: / Line 3,209: @@
 <h4><b>Week 39</b></h4>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Validation by MIC-test for  K2018010. Date: 16.09.28</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Validation by MIC-test for  K2018010. Date: 16.09.28</button>
 <div id="foo" class="panel">
 <p>The effect of the bacteriocins is validated by a MIC-test, which is made  by following the <a href="https://static.igem.org/mediawiki/2016/a/a8/T--SDU-Denmark--iGEM2016SOP0027_v01-MIC.pdf" target="_blank">SOP0027</a> step.  7.2→ 7.9.</p>
@@ Line 3,248: / Line 3,248: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Validation by MIC-test for  K2018011. Date: 16.09.28</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Validation by MIC-test for  K2018011. Date: 16.09.28</button>
 <div id="foo" class="panel">
 <p>The effect of the bacteriocins is validated by a MIC-test, which is made  by following the <a href="https://static.igem.org/mediawiki/2016/a/a8/T--SDU-Denmark--iGEM2016SOP0027_v01-MIC.pdf" target="_blank">SOP0027</a> step.  7.2→ 7.9.</p>
@@ Line 3,288: / Line 3,288: @@
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Validation by MIC-test for  K2018012. Date: 16.09.28</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Validation by MIC-test for  K2018012. Date: 16.09.28</button>
 <div id="foo" class="panel">
 <p>The effect of the bacteriocins is validated by a MIC-test, which is made  by following the <a href="https://static.igem.org/mediawiki/2016/a/a8/T--SDU-Denmark--iGEM2016SOP0027_v01-MIC.pdf" target="_blank">SOP0027</a> step.  7.2→ 7.9.</p>
@@ Line 3,326: / Line 3,326: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Validation by MIC-test for  K2018014. Date: 16.09.28</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Validation by MIC-test for  K2018014. Date: 16.09.28</button>
 <div id="foo" class="panel">
 <p>The effect of the bacteriocins is validated by a MIC-test, which is made  by following the <a href="https://static.igem.org/mediawiki/2016/a/a8/T--SDU-Denmark--iGEM2016SOP0027_v01-MIC.pdf" target="_blank">SOP0027</a> step.  7.2→ 7.9.</p>
@@ Line 3,363: / Line 3,363: @@
 </div>
-<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" height="20%" style="float:right;margin-left:15px;">Validation by MIC-test for  K2018015. Date: 16.09.28</button>
+<button class="accordion"><img src="https://static.igem.org/mediawiki/2016/e/e9/T--SDU-Denmark--minibacto.png" width="70%" style="margin:auto;">Validation by MIC-test for  K2018015. Date: 16.09.28</button>
 <div id="foo" class="panel">
 <p>The effect of the bacteriocins is validated by a MIC-test, which is made  by following the <a href="https://static.igem.org/mediawiki/2016/a/a8/T--SDU-Denmark--iGEM2016SOP0027_v01-MIC.pdf" target="_blank">SOP0027</a> step.  7.2→ 7.9.</p>

Difference between revisions of "Team:SDU-Denmark/Notebook"

Revision as of 13:05, 18 October 2016

Bacto-Aid

Notebook

Week 10

Week 13

Week 14

Week 15

Week 17

Week 18

Week 24

Week 26

Week 27

Week 28

Week 29

Week 30

Week 31

Week 32

Week 33

Week 34

Week 35

Week 36

Week 37

Week 38

Week 39