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Team:Paris Saclay/Notebook/August/2
From 2013.igem.org
Notebook : August 2
Lab work
A - Aerobic/Anaerobic regulation system
Objective : obtaining Bba_K1155003
1 - Extraction of Bba_K1155003 from DH5α
Damir, Nadia
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Tranformation from 07/30/13 works. We will extract bba_K1155003 |
Protocol : Hight copy plamid extraction
We used colony 9, 11 and 12. We mix our DNA in 50µL of H2O.
2 - Electrophoresis to check the extraction of Bba_K1155003
Damir, Nadia
| File:Ps.jpg|350px]] |
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Expected sizes :
- Bba_K1155003 : 2734 pb
Estimated concentrations :
- Clone 9 : 18ng/µL
- Clone 11 :18ng/µL
- Clone 12 : 18ng/µL
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We obtain fragment at the right size (Clone 12 starts 5 min after the others). The extraction was good. We will digest it. |
Objective : obtaining Bba_K1155007
1 - Digestion of Bba_I732017 by EcoRI/SpeI
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 don't obtain fragments at the right size. We will digest Bba_KI732017 again. |
A - Aerobic/Anaerobic regulation system / B - PCB sensor system
Objective : obtaining FNR and BphR2 proteins
1 - Transformation of BphR2, FNR and RBS-FNR in DH5α
XiaoJing
Protocol : Bacterial transformation
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