IGEM:MIT/2007/Notebook/2007-7-31
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Agenda
- Run mercury assay
- Redo live immunostaining with altered protocol
- Enter working parts into BioBricks Registry
- Find constitutive promoter and perform 3A assembly with a RBS and CPX
- Redo transformation of BL21 w/ mer.I13500 in 1AT3
Mercury Assay
CPX Assay
Varied cell concentrations: (OD)
- .1
- .3
- .5
- 1
CPX Immunostaining
- Induced cells at 0.1 OD, incubate for 1 hour
- 10 samples total: 2 each for 0(- control), 10^-8, 10^-7, 10^-6, 10^-5 concentration AHL
- Changed secondary Goat Anti-Mouse antibody concentration to (1:200)
Results
-
AHL 5Ai
-
AHL 5Aii
-
AHL 5Bi
-
AHL 5Bii
-
AHL 6Ai
-
AHL 6Aii
-
AHL 6Bi
-
AHL 7Ai
-
AHL 7Aii
-
AHL 7Aiii
-
AHL 7Bi
-
AHL 7Bii
-
AHL 8Ai
-
AHL 8Aii
-
AHL 8Bi
-
AHL 8Bii
-
Control Ai
-
Control Aii
-
Control Bi
-
Control Bii
Transformation of mer.I13500 in BL21
- Nothing grew on any of the plates (not even positive control with pUC18)
- Perhaps cells not competent?
- We should redo tomorrow