Team:TU Delft/22 September 2010 content

From 2010.igem.org

(Difference between revisions)
(New page: ==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 as...)
(→Emulsifier)
 
(4 intermediate revisions not shown)
Line 34: Line 34:
|
|
|}
|}
 +
 +
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.

Latest revision as of 12:35, 1 October 2010

Contents

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.

Retrieved from "http://2010.igem.org/Team:TU_Delft/22_September_2010_content"