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Team:Paris Saclay/Notebook/August/19
From 2013.igem.org
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* H2O : 13µL | * H2O : 13µL | ||
* Ligase : 1µL | * Ligase : 1µL | ||
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| + | ====3 - Transformation of ligation of NarK, NarG or NirB with RBS-LacZ-Term or RBS-AmilCP-Term in PSB1C3==== | ||
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| + | XiaoJing | ||
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| + | Protocol : [[Team:Paris_Saclay/Protocols/bacterial transformation|Bacterial transformation]] | ||
Revision as of 19:45, 11 September 2013
Notebook : August 19
Lab work
A - Aerobic/Anaerobic regulation system
Objective : to obtain ...
1 - Gel purification of the digestion of Bba_K1155003 and Bba_K1155007 by XBaI/PstI
Nadia
Protocol : Gel purification
Nanodrop :
- RBS-LacZ-Term : 59.6ng/µL
- RBS-AmilCP-Term : 21.3ng/µL
|
Concentrations are very low so we try to precipitate the DNA again. |
2 - EtOH precipitation of RBS-LacZ-Term and RBS-AmilCP-Term
Nadia, XiaoJing
Protocol : EtOH precipitation Protocole ou à rédiger ici ?????????
We used 20µL of DNA.
Nanodrop :
- RBS-LacZ-Term : 38.4ng/µL
- RBS-AmilCP-Term : 28.7ng/µL
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CONCLUSION |
3 - Ligation of NarK, NarG or NirB with RBS-LacZ-Term or RBS-AmilCP-Term in PSB1C3
XiaoJing
Used quantities :
- NirB and RBS-LacZ-Term :
- NirB : 3µL
- RBS-LacZ-Term : 1µL
- Buffer : 2µL
- H2O : 13µL
- Ligase : 1µL
- NirB and RBS-AmilCP-Term :
- NirB : 3µL
- RBS-AmilCP-Term : 1µL
- Buffer : 2µL
- H2O : 13µL
- Ligase : 1µL
- NarG and RBS-LacZ-Term :
- NarG : 3µL
- RBS-LacZ-Term : 1µL
- Buffer : 2µL
- H2O : 13µL
- Ligase : 1µL
- NarG and RBS-AmilCP-Term :
- NarG : 3µL
- RBS-AmilCP-Term : 1µL
- Buffer : 2µL
- H2O : 13µL
- Ligase : 1µL
- NarK and RBS-LacZ-Term :
- Nar K : 3µL
- RBS-LacZ-Term : 1µL
- Buffer : 2µL
- H2O : 13µL
- Ligase : 1µL
- NarK and RBS-AmilCP-Term :
- NarK : 3µL
- RBS-AmilCP-Term : 1µL
- Buffer : 2µL
- H2O : 13µL
- Ligase : 1µL
3 - Transformation of ligation of NarK, NarG or NirB with RBS-LacZ-Term or RBS-AmilCP-Term in PSB1C3
XiaoJing
Protocol : Bacterial transformation
A - Aerobic/Anaerobic regulation system / B - PCB sensor system
Objective : obtaining FNR and BphR2 proteins (Gibson assembly)
1 - Electrophoresis gel of PCR products : BphR2 Part I
Damir
| [[]] |
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Expected size :
- BphR2 Part I : 178 kb
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We obtain a frangment at the right size. We can purify it. |
2 - Gel purification of PCR products : BphR2 Part I
Damir
Protocol : Gel purification
Nanodrop :
- BphR2 : 57.2ng/µL
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CONCLUSION |
Meeting with Evry
