Team:Paris Saclay/Notebook/July/22
From 2013.igem.org
Notebook : July 22
Lab work
A - Aerobic/Anaerobic regulation system
Objective : obtaining Bba_K1155003, Bba_K1155007
1 -Digestion of Bba_I732017 by EcoRi/SpeI, Bba_K592009 by SpeI/PstI, Bba_B0015 and Bba_B0017 by PstI/XbaI
Anaïs
Used quantities :
- Bba_I732017 :
- Buffer FD : 3µL
- Bba_I732017 : 15µL
- EcoRI FD : 2µL
- SpeI FD : 2µL
- H2O : 8µL
- Bba_K502009 :
- Buffer FD : 3µL
- Bba_K502009 : 15µL
- PstI FD : 2µL
- SpeI FD : 2µL
- H2O : 8µL
- Bba_B0015 :
- Buffer tango : 3µL
- Bba_B0015 : 15µL
- PstI FD : 1µL
- XbaI FD : 2µL
- H2O : 9µL
- Bba_B0017 :
- Buffer tango : 3µL
- Bba_B0017 : 15µL
- PstI FD : 1µL
- XbaI FD : 2µL
- H2O : 9µL
We let the digestion 1h30 at 37°C and then we put it 30 min in the incubator at 37°C because the temperature of the bain-marie decrease at 31°C after 1h30.
2 - Denaturation of EcoRI/SpeI, SpeI/PstI and PstI/XbaI used for the digestion of Bba_I732017, Bba_K502009, Bba_B0015 and Bba_B0017
Zhou
Protocol : Ethanol precipitation
We used 30µL of DNA.