Team:TU Delft/22 September 2010 content

From 2010.igem.org

(Difference between revisions)
(→Emulsifier)
(→Emulsifier assay)
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The samples were immediately subjected to the emulsifier assay with Sudan II.
The samples were immediately subjected to the emulsifier assay with Sudan II.
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T=0
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T=0 (Tris Blank + Sudan II = 0.282)
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T=2 (Tris Blank + Sudan II = 0.322)
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{|
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|'''#'''
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|'''Culture'''
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|'''Sudan II'''
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|'''Absorption 493 nm'''
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|1
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|1.408
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|1.601
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T=4 (Tris Blank + Sudan II = 0.280)
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{|
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|'''#'''
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|'''Culture'''
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|'''Sudan II'''
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|'''Absorption 493 nm'''
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|1
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|1.441
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|1.716
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|1.656
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|5
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|3
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|1.460
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|6
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|3
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|1.759
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Revision as of 12:19, 24 September 2010

Emulsifier

The K12 cultures were grown over night on M9. We tried to do 2 experiments today. One for getting samples for a protein gel and another one for samples for the emulsifier assay.

Protein gel

One 50 mL tube with 10 mL M9 + Glucose was inocculated with K12 that was transformed with 206. Once the OD600 was 0.6 the protein production was induced by adding IPTG to a final concentration of 1 mM. Samples were taken every hour, for 4 hours and over night.

# Time point OD600
1 0 0.6
2 1 1.0
3 2 1.16
4 3 1.17
5 4
6 0/n

0.5 mL samples were taken, spinned down, and the sup and pellet were stored at -20.

Emulsifier assay

The experiment was done in triplo. 50 mL tubes were filled with 10 mL M9 + glucose and inocculated. Once OD600 was 0.6 the protein production was induced by adding IPTG to a final concentration of 1 mM.

1 mL samples were taken at T=0, 2 and 4.

The samples were immediately subjected to the emulsifier assay with Sudan II.

T=0 (Tris Blank + Sudan II = 0.282)

# Culture Sudan II Absorption 493 nm
1 1 - 0.981
2 1 + 1.211
3 2 - 0.985
4 2 + 1.266
5 3 - 0.985
6 3 - 1.243

T=2 (Tris Blank + Sudan II = 0.322)

# Culture Sudan II Absorption 493 nm
1 1 - 1.408
2 1 + 1.745
3 2 - 1.417
4 2 + 1.736
5 3 - 1.404
6 3 - 1.601

T=4 (Tris Blank + Sudan II = 0.280)

# Culture Sudan II Absorption 493 nm
1 1 - 1.441
2 1 + 1.716
3 2 - 1.443
4 2 + 1.656
5 3 - 1.460
6 3 - 1.759
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