Team:Paris Saclay/Notebook/August/6
From 2013.igem.org
Notebook : August 6
Lab work
A - Aerobic/Anaerobic regulation system
Objective : obtaining Bba_K1155004, Bba_K1155005, Bba_K1155006
1 - Electrophoresis to check the Colony PCR products : Bba_K1155004, Bba_K1155005, Bba_K1155006
XiaoJing, Damir, Anaïs
- Bba_K1155004 :
[[]] |
|
- Bba_K1155005 :
[[]] |
|
- Bba_K1155006 :
[[]] |
|
Expected size :
- NarK, NarG, NirB : 500 bp
We obtain fragments at the good size for all the colony. We will make a culture of DH5α with Bba_K1155004, Bba_K1155005 and Bba_K1155006. We will also sequence our plasmids. |
2 - Culture of DH5α with Bba_K1155004, Bba_K1155005 and Bba_K1155006
Xiaoing, Anaïs
Used quantities :
- LB : 5mL
- Chloramphénicol (1000X, 20µg/mL) : 5µL
- Bba_K1155004, Bba_K1155005 and Bba_K1155006 : 25µL ?????????
We let the incubation over night at 37°C at 180 RPM.
We used colonies number 6, 7 and 8 for each promotor.
Objective : obtaining Bba_K1155007
1 - Electroelution of Bba_I732017 digested by ecoRI/SpeI
Nadia
Protocol : Electroelution
The electroelution was good. We will ligate RBS-LacZ with Term and PSB1C3. |
B - PCB sensing system
Objective : obtaining Bba_K1155002
1 - Sequence analysis for Bba_K1155002 in clones 4, 17 and 22
IMAGE LOGICIEL
The sequence is good for each clone. We obtain a new biobrick : Bba_K1155002. |