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− | {{UBonn_HBRS/header|title=Lab Notebook 2016|Project=active}} | + | {{UBonn_HBRS/header|title=Lab Notebook 2016|Project=active|class-main=col-sm-12}} |
− | 10KW (7.3-13.3) | + | <html><div class="panel notebook-entry"></html> |
− | + | ==10KW (7.3-13.3)== | |
transformation of genes bought at genescript: XynA, XynB, LipA, EstC2, CpCel9, CpCel5c, EngB, CelA, BsCel5, Bpul | transformation of genes bought at genescript: XynA, XynB, LipA, EstC2, CpCel9, CpCel5c, EngB, CelA, BsCel5, Bpul | ||
− | and Linkers: nprb, SacB | + | and Linkers: nprb, SacB <br /> |
inoculation of liquid cultures and restriction Analysi of linkers (SacB and nprb). | inoculation of liquid cultures and restriction Analysi of linkers (SacB and nprb). | ||
− | repetition of the transformation for XynB, EngB and linkers. | + | repetition of the transformation for XynB, EngB and linkers. <br /> |
Subsequently inoculation and miniprep of the samples. Preparation of glycerol stock. | Subsequently inoculation and miniprep of the samples. Preparation of glycerol stock. | ||
restriction analysis of the genes. | restriction analysis of the genes. | ||
+ | <html></div></html> | ||
− | |||
+ | <html><div class="panel notebook-entry"></html> | ||
+ | ==11KW (14.3-20.3)== | ||
Repetion of restriction analysis from the genes KW10 and gel clean ups of the linkers. | Repetion of restriction analysis from the genes KW10 and gel clean ups of the linkers. | ||
− | + | Deinking: <br /> | |
+ | * Measurement of inkjet absorbance spectrum | ||
+ | * filtration-based deinking setup | ||
+ | <html></div></html> | ||
+ | |||
+ | <html><div class="panel notebook-entry"></html> | ||
+ | ==12KW (21.3-27.3)== | ||
repetition of restriction analysis of all genes and gel clean ups of the genes. | repetition of restriction analysis of all genes and gel clean ups of the genes. | ||
− | + | Deinking: <br /> | |
+ | inkjet ink absorbance spectrum and pulp preparation | ||
+ | <html></div></html> | ||
+ | |||
+ | <html><div class="panel notebook-entry"></html> | ||
+ | ==13KW (28.3-3.4)== | ||
Restrictions and gel clean ups of all genes genes for a new gel clean up, because the ones of KW13 did not works. Ligation of genes from the gel clean ups into C3 and inoculation of cultures and minipreps. | Restrictions and gel clean ups of all genes genes for a new gel clean up, because the ones of KW13 did not works. Ligation of genes from the gel clean ups into C3 and inoculation of cultures and minipreps. | ||
(1.04.16 :) | (1.04.16 :) | ||
− | + | → successful ligation of | |
− | XynA | + | * XynA |
− | CelA | + | * CelA |
− | LipA | + | * LipA |
− | Bpul | + | * Bpul |
− | CpCel5c | + | * CpCel5c |
− | BsCel5 | + | * BsCel5 |
− | EstC2 | + | * EstC2 |
− | EngB in C3 | + | * EngB in C3 |
+ | |||
+ | * nprb + CpCel9 | ||
+ | * nprb + BsCel5 in C3 | ||
+ | |||
+ | |||
+ | Deinking: <br /> | ||
+ | establishing the set up and starting experiments with different conc of NaOH and Na2SiO3 | ||
+ | <html></div></html> | ||
+ | |||
+ | |||
+ | <html><div class="panel notebook-entry"></html> | ||
+ | ==14KW (4.4-10.4)== | ||
+ | Deinking: <br /> | ||
+ | Experiments with different concentrations of surfactant oleic acid | ||
+ | <html></div></html> | ||
+ | |||
+ | |||
+ | <html><div class="panel notebook-entry"></html> | ||
+ | ==15KW (11.4-17.4)== | ||
+ | Deinking: <br /> | ||
+ | experiments with different incubation time, vortexing time, temperature | ||
+ | <html></div></html> | ||
+ | |||
+ | |||
+ | <html><div class="panel notebook-entry"></html> | ||
+ | ==16KW (18.4-24.4)== | ||
+ | Deinking: <br /> | ||
+ | tests with solvents to dissolve ink and fibers | ||
+ | <html></div></html> | ||
+ | |||
+ | |||
+ | <html><div class="panel notebook-entry"></html> | ||
+ | ==17KW (25.4-1.5)== | ||
+ | Deinking: <br /> | ||
+ | grayscale tests, solvent tests | ||
+ | <html></div></html> | ||
+ | |||
+ | |||
+ | <html><div class="panel notebook-entry"></html> | ||
+ | ==18KW (2.5-8.5)== | ||
+ | Deinking: <br /> | ||
+ | pulp preparation | ||
+ | <html></div></html> | ||
+ | |||
+ | |||
+ | <html><div class="panel notebook-entry"></html> | ||
+ | ==19KW (9.5-15.5)== | ||
+ | Deinking: <br /> | ||
+ | establishing water and standard chemical controls | ||
+ | <html></div></html> | ||
+ | |||
− | + | <html><div class="panel notebook-entry"></html> | |
− | + | ==20KW (16.5-22.5)== | |
+ | Deinking: <br /> | ||
+ | establishing water and standard chemical controls | ||
+ | <html></div></html> | ||
− | |||
+ | <html><div class="panel notebook-entry"></html> | ||
+ | ==21KW (23.5-29.5)== | ||
Restriction analysis of genes from the 13KW and mini prep from samples of the 13KW. | Restriction analysis of genes from the 13KW and mini prep from samples of the 13KW. | ||
(28.05.16) | (28.05.16) | ||
successful ligation of: | successful ligation of: | ||
− | nprb and sacB in C3 | + | * nprb and sacB in C3 |
− | nprb + EstC2 in C3 | + | * nprb + EstC2 in C3 |
− | + | Deinking: <br /> | |
+ | establishing oleic acid and oleic acid + citrate buffer controls | ||
+ | <html></div></html> | ||
+ | |||
+ | <html><div class="panel notebook-entry"></html> | ||
+ | ==22KW (30.5-5.6)== | ||
Restriction analysis of samples of the 21KW | Restriction analysis of samples of the 21KW | ||
− | + | Deinking: <br /> | |
+ | discovery: paper discs with CB turn red, tests to find the reason | ||
+ | <html></div></html> | ||
+ | |||
+ | <html><div class="panel notebook-entry"></html> | ||
+ | ==23KW (6.6-12.6)== | ||
Inoculation of all genes from glycerol stocks and miniprep. Restriction analysis and gel clean up of the genes, ligation of the missing genes with C3. | Inoculation of all genes from glycerol stocks and miniprep. Restriction analysis and gel clean up of the genes, ligation of the missing genes with C3. | ||
− | 24KW (13.6-19.6) | + | Deinking: <br /> |
+ | tests with oleic acid and oleic acid + cb controls | ||
+ | <html></div></html> | ||
+ | |||
+ | |||
+ | <html><div class="panel notebook-entry"></html> | ||
+ | ==24KW (13.6-19.6)== | ||
+ | Restriction of bbMCS and gel clean ups. <br /> | ||
+ | Ligations of genes in C5 and C3. <br /> | ||
+ | Ligation of genes and tags in C5. <br /> | ||
− | |||
− | |||
− | |||
Minipreps of all of the samples. | Minipreps of all of the samples. | ||
− | + | Deinking: <br /> | |
+ | tests with shaking during incubation and different incubation time | ||
+ | <html></div></html> | ||
+ | |||
+ | <html><div class="panel notebook-entry"></html> | ||
+ | ==25KW (20.6-26.6)== | ||
Restriction analysis of the constructs | Restriction analysis of the constructs | ||
+ | |||
Repetition of ligation of genes in C5 and transformation and preps. | Repetition of ligation of genes in C5 and transformation and preps. | ||
− | 26KW (27.6 - 3.7) | + | Deinking: <br /> |
+ | rotation evaporator concentrating ink | ||
+ | <html></div></html> | ||
+ | |||
+ | |||
+ | <html><div class="panel notebook-entry"></html> | ||
+ | ==26KW (27.6 - 3.7)== | ||
Restriction analysis of the mini preps. | Restriction analysis of the mini preps. | ||
+ | <html></div></html> | ||
− | 28KW (11.7- | + | |
+ | <html><div class="panel notebook-entry"></html> | ||
+ | ==28KW (11.7-17.7)== | ||
Repetion of Ligations of genes and C5 | Repetion of Ligations of genes and C5 | ||
− | + | ''B.subtilis'': | |
+ | preparing electrocomp cells and transformations | ||
+ | |||
+ | Deinking: <br /> | ||
+ | flotation-based deinking, pulp preparation | ||
+ | <html></div></html> | ||
+ | |||
+ | |||
+ | <html><div class="panel notebook-entry"></html> | ||
+ | ==29KW (18.7-24.7)== | ||
+ | Deinking: <br /> | ||
+ | separation of ink from fibers in organic phase of big scale deinking, flotation-based deinking, foam collection, defoaming | ||
+ | <html></div></html> | ||
+ | |||
+ | |||
+ | <html><div class="panel notebook-entry"></html> | ||
+ | ==30KW (25.7-31.7)== | ||
+ | ''B.subtilis'': <br /> | ||
+ | preparing electrocomp cells and transformations | ||
+ | preparing chemocompetent B. subtilis and transformations | ||
+ | |||
+ | Deinking: <br /> | ||
+ | tests with toluene flotation-based deinking | ||
+ | <html></div></html> | ||
+ | |||
− | Ligation of Fragment synthesized at IDT and C3 | + | <html><div class="panel notebook-entry"></html> |
− | Ligation of genes in C3 and genes + tags + C5 | + | ==31KW (1.8-7.8)== |
+ | Ligation of Fragment synthesized at IDT and C3. <br /> | ||
+ | Ligation of genes in C3 and genes + tags + C5. <br /> | ||
transformation, and inoculation of liquid cultures of the above. | transformation, and inoculation of liquid cultures of the above. | ||
− | + | ''B.subtilis'': <br /> | |
+ | preparing of chemocompetent cells and transformations | ||
+ | <html></div></html> | ||
− | |||
− | |||
− | |||
− | + | <html><div class="panel notebook-entry"></html> | |
− | Transformation of | + | ==32KW (8.8 - 14.8)== |
+ | Ligation of IDT and C3. <br /> | ||
+ | Transformation and inoculation. <br /> | ||
+ | Minipreps of samples and restriction analysis. | ||
− | + | Ligations of genes plus tags in bb_MCS. <br /> | |
+ | Transformation of these samples. | ||
− | Inoculation and miniprep | + | ''B.subtilis'': <br /> |
− | Repetion of ligation of IDT and C3 | + | preparing of electrocomp and chemocompetent cells and transformations |
− | Repetion of ligation of bb + nprb + genes | + | <html></div></html> |
− | Midi prep of SacB, bb and nprb | + | |
− | Ligation of Genes + C5 and genes plus tags in C5. | + | |
− | Transformation and inoculation of the samples. | + | <html><div class="panel notebook-entry"></html> |
+ | ==33KW (15.8 - 21.8)== | ||
+ | Inoculation and miniprep. <br /> | ||
+ | Repetion of ligation of IDT and C3. <br /> | ||
+ | Repetion of ligation of bb + nprb + genes. <br /> | ||
+ | Midi prep of SacB, bb and nprb. <br /> | ||
+ | Ligation of Genes + C5 and genes plus tags in C5. <br /> | ||
+ | Transformation and inoculation of the samples. <br /> | ||
mini preps | mini preps | ||
− | + | Deinking: <br /> | |
+ | creation of flat paper-like discs | ||
+ | <html></div></html> | ||
+ | |||
+ | <html><div class="panel notebook-entry"></html> | ||
+ | ==34KW (22.8 - 28.8)== | ||
Restrictions of mini prep and repetition of mini prep | Restrictions of mini prep and repetition of mini prep | ||
− | {{UBonn_HBRS/footer}} | + | |
+ | ''B.subtilis'': <br/> | ||
+ | preparing of chemocompetent cells and transformations | ||
+ | |||
+ | Deinking: <br /> | ||
+ | grayscale tests with officer scanner and odisey scanner | ||
+ | <html></div></html> | ||
+ | |||
+ | |||
+ | <html><div class="panel notebook-entry"></html> | ||
+ | ==35KW (29.9 - 5.9)== | ||
+ | ''B.subtilis'': <br/> | ||
+ | transformations of bb | ||
+ | <html></div></html> | ||
+ | |||
+ | |||
+ | <html><div class="panel notebook-entry"></html> | ||
+ | ==36KW (5.9 - 11.9)== | ||
+ | Deinking: <br /> | ||
+ | tests of water controls | ||
+ | <html></div></html> | ||
+ | |||
+ | |||
+ | <html><div class="panel notebook-entry"></html> | ||
+ | ==37KW (12.9 - 18.9)== | ||
+ | ''B.subtilis'': <br/> | ||
+ | transformations of bb | ||
+ | |||
+ | Deinking: <br /> | ||
+ | scanning tests, different conditions tests | ||
+ | <html></div></html> | ||
+ | |||
+ | |||
+ | <html><div class="panel notebook-entry"></html> | ||
+ | ==38KW (19.9 - 25.9)== | ||
+ | ''B.subtilis'': <br/> | ||
+ | preparing chemocompetent cells and transformations | ||
+ | <html></div></html> | ||
+ | |||
+ | |||
+ | <html><div class="panel notebook-entry"></html> | ||
+ | ==39KW (26.9 - 2.10)== | ||
+ | Deinking: <br /> | ||
+ | establishing water and standard controls into setup with kitchen mixer | ||
+ | <html></div></html> | ||
+ | |||
+ | |||
+ | <html><div class="panel notebook-entry"></html> | ||
+ | ==40KW (3.10 - 9.10)== | ||
+ | Deinking: <br /> | ||
+ | xylanase started to test + cb | ||
+ | <html></div></html> | ||
+ | |||
+ | |||
+ | <html><div class="panel notebook-entry"></html> | ||
+ | ==41KW (10.10 - 16.10)== | ||
+ | Deinking: <br /> | ||
+ | xyl tests + cb | ||
+ | <html></div></html> | ||
+ | {{UBonn_HBRS/footer|class-sideNav=hidden}} |
Latest revision as of 21:47, 19 October 2016
Lab Notebook 2016
Contents
- 1 10KW (7.3-13.3)
- 2 11KW (14.3-20.3)
- 3 12KW (21.3-27.3)
- 4 13KW (28.3-3.4)
- 5 14KW (4.4-10.4)
- 6 15KW (11.4-17.4)
- 7 16KW (18.4-24.4)
- 8 17KW (25.4-1.5)
- 9 18KW (2.5-8.5)
- 10 19KW (9.5-15.5)
- 11 20KW (16.5-22.5)
- 12 21KW (23.5-29.5)
- 13 22KW (30.5-5.6)
- 14 23KW (6.6-12.6)
- 15 24KW (13.6-19.6)
- 16 25KW (20.6-26.6)
- 17 26KW (27.6 - 3.7)
- 18 28KW (11.7-17.7)
- 19 29KW (18.7-24.7)
- 20 30KW (25.7-31.7)
- 21 31KW (1.8-7.8)
- 22 32KW (8.8 - 14.8)
- 23 33KW (15.8 - 21.8)
- 24 34KW (22.8 - 28.8)
- 25 35KW (29.9 - 5.9)
- 26 36KW (5.9 - 11.9)
- 27 37KW (12.9 - 18.9)
- 28 38KW (19.9 - 25.9)
- 29 39KW (26.9 - 2.10)
- 30 40KW (3.10 - 9.10)
- 31 41KW (10.10 - 16.10)
10KW (7.3-13.3)
transformation of genes bought at genescript: XynA, XynB, LipA, EstC2, CpCel9, CpCel5c, EngB, CelA, BsCel5, Bpul
and Linkers: nprb, SacB
inoculation of liquid cultures and restriction Analysi of linkers (SacB and nprb).
repetition of the transformation for XynB, EngB and linkers.
Subsequently inoculation and miniprep of the samples. Preparation of glycerol stock.
restriction analysis of the genes.
11KW (14.3-20.3)
Repetion of restriction analysis from the genes KW10 and gel clean ups of the linkers.
Deinking:
- Measurement of inkjet absorbance spectrum
- filtration-based deinking setup
12KW (21.3-27.3)
repetition of restriction analysis of all genes and gel clean ups of the genes.
Deinking:
inkjet ink absorbance spectrum and pulp preparation
13KW (28.3-3.4)
Restrictions and gel clean ups of all genes genes for a new gel clean up, because the ones of KW13 did not works. Ligation of genes from the gel clean ups into C3 and inoculation of cultures and minipreps. (1.04.16 :) → successful ligation of
- XynA
- CelA
- LipA
- Bpul
- CpCel5c
- BsCel5
- EstC2
- EngB in C3
- nprb + CpCel9
- nprb + BsCel5 in C3
Deinking:
establishing the set up and starting experiments with different conc of NaOH and Na2SiO3
14KW (4.4-10.4)
Deinking:
Experiments with different concentrations of surfactant oleic acid
15KW (11.4-17.4)
Deinking:
experiments with different incubation time, vortexing time, temperature
16KW (18.4-24.4)
Deinking:
tests with solvents to dissolve ink and fibers
17KW (25.4-1.5)
Deinking:
grayscale tests, solvent tests
18KW (2.5-8.5)
Deinking:
pulp preparation
19KW (9.5-15.5)
Deinking:
establishing water and standard chemical controls
20KW (16.5-22.5)
Deinking:
establishing water and standard chemical controls
21KW (23.5-29.5)
Restriction analysis of genes from the 13KW and mini prep from samples of the 13KW. (28.05.16) successful ligation of:
- nprb and sacB in C3
- nprb + EstC2 in C3
Deinking:
establishing oleic acid and oleic acid + citrate buffer controls
22KW (30.5-5.6)
Restriction analysis of samples of the 21KW
Deinking:
discovery: paper discs with CB turn red, tests to find the reason
23KW (6.6-12.6)
Inoculation of all genes from glycerol stocks and miniprep. Restriction analysis and gel clean up of the genes, ligation of the missing genes with C3.
Deinking:
tests with oleic acid and oleic acid + cb controls
24KW (13.6-19.6)
Restriction of bbMCS and gel clean ups.
Ligations of genes in C5 and C3.
Ligation of genes and tags in C5.
Minipreps of all of the samples.
Deinking:
tests with shaking during incubation and different incubation time
25KW (20.6-26.6)
Restriction analysis of the constructs
Repetition of ligation of genes in C5 and transformation and preps.
Deinking:
rotation evaporator concentrating ink
26KW (27.6 - 3.7)
Restriction analysis of the mini preps.
28KW (11.7-17.7)
Repetion of Ligations of genes and C5
B.subtilis: preparing electrocomp cells and transformations
Deinking:
flotation-based deinking, pulp preparation
29KW (18.7-24.7)
Deinking:
separation of ink from fibers in organic phase of big scale deinking, flotation-based deinking, foam collection, defoaming
30KW (25.7-31.7)
B.subtilis:
preparing electrocomp cells and transformations
preparing chemocompetent B. subtilis and transformations
Deinking:
tests with toluene flotation-based deinking
31KW (1.8-7.8)
Ligation of Fragment synthesized at IDT and C3.
Ligation of genes in C3 and genes + tags + C5.
transformation, and inoculation of liquid cultures of the above.
B.subtilis:
preparing of chemocompetent cells and transformations
32KW (8.8 - 14.8)
Ligation of IDT and C3.
Transformation and inoculation.
Minipreps of samples and restriction analysis.
Ligations of genes plus tags in bb_MCS.
Transformation of these samples.
B.subtilis:
preparing of electrocomp and chemocompetent cells and transformations
33KW (15.8 - 21.8)
Inoculation and miniprep.
Repetion of ligation of IDT and C3.
Repetion of ligation of bb + nprb + genes.
Midi prep of SacB, bb and nprb.
Ligation of Genes + C5 and genes plus tags in C5.
Transformation and inoculation of the samples.
mini preps
Deinking:
creation of flat paper-like discs
34KW (22.8 - 28.8)
Restrictions of mini prep and repetition of mini prep
B.subtilis:
preparing of chemocompetent cells and transformations
Deinking:
grayscale tests with officer scanner and odisey scanner
35KW (29.9 - 5.9)
B.subtilis:
transformations of bb
36KW (5.9 - 11.9)
Deinking:
tests of water controls
37KW (12.9 - 18.9)
B.subtilis:
transformations of bb
Deinking:
scanning tests, different conditions tests
38KW (19.9 - 25.9)
B.subtilis:
preparing chemocompetent cells and transformations
39KW (26.9 - 2.10)
Deinking:
establishing water and standard controls into setup with kitchen mixer
40KW (3.10 - 9.10)
Deinking:
xylanase started to test + cb
41KW (10.10 - 16.10)
Deinking:
xyl tests + cb