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Team:Paris Saclay/Notebook/August/5
From 2013.igem.org
Notebook : August 5
Lab work
A - Aerobic/Anaerobic regulation system
Objective : obtaining Bba_K1155003
1 - Digestion of Bba_K1155003 by EcoRI/PstI to check the ligation between RBS-Amil CP,Term and PSB1C3
Nadia, XiaoJing
Used quantities :
- Bba_K1155003 : 5µL
- EcoRI FD : 1µL
- PstI FD : 1µL
- Buffer FD : 3µL
- H2O : 20µL
We let the digestion at 37°C during 15 minutes.
2 - Electrophoresis of the digestion of Bba_K1155003
Damir
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Expected sizes :
- RBS_Amil CP-Term :
- PSB1C3 :
Objective : obtaining Bba_K1155007
1 - Digestion of Bba_I732017 by EcoRI/SpeI
Abdou, Damir, Nadia,
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
Damir, Nadia
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Expected sizes :
- RBS-LacZ : 3093 bp
- PSB1A2 : 2079 bp
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We obtain fragments at the right size. We will make an elector-elution to extract RBS-LacZ. |
Objective : obtaining Bba_K1155004, Bba_K1155005, Bba_K1155006
1 - Colony PCR of Bba_K1155004, Bba_K115005, Bba_K1155006 in DH5α
Damir, Nadia, XiaoJing
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Transformation of 07/31/13 works. We will do a PCR Colony. |
COLONIES PIQUEES DANS 10µL d'eau par Tube !!!!!!!!!!!!!
Used quantities :
- DNA : 2µL
- Mix : (it was divided in 25 tubes for each promotor with 23µL of mix in each tube)
- Oligo 44 : 3.5µL
- Oligo 45 : 3.5µL
- Buffer Dream Taq : 70µL
- dNTP : 28µL
- Dream Taq : 5µL
- H2O : 590µL
PCR Program :
