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From 2010.igem.org

Contents 1 Emulsifier 1.1 Protein gel 1.2 Emulsifier assay 2 Alkane degradation

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

Alkane degradation

Pieter was so kind to isolate the 028 plasmids from the 5 positives of yesterday. Cellbanks were also made, and competent K12 cells were transformed.