Team:TU Delft/22 September 2010 content
From 2010.igem.org
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 |