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Team:Paris Saclay/Notebook/July/31
From 2013.igem.org
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| Line 14: | Line 14: | ||
Used quantities : | Used quantities : | ||
| - | * PSB1C3 : | + | * PSB1C3 : 16µL |
| - | * Buffer FD : | + | * Buffer FD : 2µL |
| - | * EcoRI FD : | + | * EcoRI FD : 1µL |
| - | * PstI FD : | + | * PstI FD : 1µL |
We let our digestion 1h30 at 37°C. | We let our digestion 1h30 at 37°C. | ||
Revision as of 14:42, 27 September 2013
Notebook : July 31
Lab work
A - Aerobic/Anaerobic regulation system
Objective : obtaining Bba_K1155004, Bba_K1155005, Bba_K155006
1 -Digestion of PSB1C3 by EcoRI/PstI
Abdou
Used quantities :
- PSB1C3 : 16µL
- Buffer FD : 2µL
- EcoRI FD : 1µL
- PstI FD : 1µL
We let our digestion 1h30 at 37°C.
2 -Denaturation of EcoRI/PstI used for the digestion of PSB1C3
Abdou
Protocol : Ethanol precipitation
3 -Ligation of NarK, NarG, NirB in PSB1C3
XiaoJing
Used quantities :
- Buffer : 2µL
- NarK, NarG, NirB : 1µL
- PSB1C3 : 2µL
- Ligase : 1µL
- H2O : 14µL
4 -Electrophoresis to check the ligation of NarK, NarG, NirB in PSB1C3
XiaoJing
| [[]] |
|
Expected sizes :
- NarK : ...
- Nar G : ...
- Nir B : ...
|
We obtain fragments at the right size. We will transform them in DH5α. |
5 -Transformation of Bba_K1155004, Bba_K1155005, Bba_K1155006
Anaïs
Protocol : Bacterial transformation
Objective : obtaining Bba_K1155007
1 - Digestion of Bba_I732017 by EcoRI/SpeI
XiaoJing
Used quantities :
- Bba_I732017 : 41µL
- Buffer FD : 5µL
- EcoRI : 2µL
- SpeI : 2µL
We let the digestion at 1h30 at 37°C.
2 -Electrophoresis to check the digestion of Bba_I732017 by EcoRI/SpeI
XiaoJing
| [[]] |
|
Expected sizes :
- RBS-LacZ : 3093 bp
- PSB1A2 : 2079 bp
|
We obtain fragments at the right size. We will purify it. |
3 - Gel purification of digestion of RBS-LacZ
Xavier
Protocol : Gel purification
|
We lost our fragments. We will do the digestion again. |
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