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Team:Paris Saclay/Notebook/August/22
From 2013.igem.org
Contents |
Notebook : August 22
Lab work
A - Aerobic/Anaerobic regulation system
Objective : characterize Bba_K1155000, Bba_K1155004, Bba_K1155005, Bba_K1155006
1 - Plasmid extraction of Bba_K1155000 from DH5α
Nguyen
Protocol : Plasmid extraction
Nanodrop :
- Bba_K1155000 : 175ng/µL
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CONCLUSION !!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!!! |
2 - Digestion of Bba_K1155000 by SpeI
Nguyen
Used quantities :
- Bba_K1155000 : 10µL
- Buffer FD : 2µL
- Spe I : 2µL
- H2O : 6 µL
We let digestions at 37°C during 10 minutes ??????
QU'EN A T'ON FAIT PAR LA SUITE ?????????
((((((((((((((((((====Objective : obtaining Pfnr, NarK, NarG or NirB and RBS-LacZ-Term or RBS-AmilCP-Term in PSB3K3====
1 - Gel purification of the digestion of Bba_J04450 by EcoRI/PstI
XiaoJing
Protocol : Gel purification
Nanodrop :
- PSB3K3 : 4ng/µL
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The Nanodrop gives us a very few quantity of PSB3K3 so we decided to check it with a first electrophoresis. |
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Expect sizes :
- PSB3K3 : 2750 bp
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We can see anything with the fisrt electrophoresis that's why we made an EtOH precipitation. |
Protocol : EtOH precipitation
We used 34µL of DNA. ))))))))))))))))))))))
A - Aerobic/Anaerobic regulation system / B - PCB sensor system
Objective : obtaining BphR2 protein
1 - PCR of BphR2 Part I
Damir
Used quantities :
- Oligo 54F : 2µL
- Oligo 55R : 2µL
- DNA : 1µL
- Buffer Phusion : 10µL
- dNTP : 1µL
- Phusion : 1µL
- DMS9 : 2µL ??????????????????????????????????
- H2O : 31µL
PCR program :
2 - Electrophoresis of PCR product : BphR2 Part I
Damir
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Expected sizes : Bphr2 Part I : 178 kb
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We obtain fragments at the right size. We can purify it. |
3 - Gel purification of PCR product : BphR2 Part I
Damir
Protocol : Gel purification
Nanodrop :
- RBS-BphR2 Part I, tube 1 : 42ng/µL
- RBS-BphR2 Part I, tube 2 : 75ng/µL
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CONCLUSION !!!!!!!!!!!!!!!!!!!!!!!!! |
