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Team:Paris Saclay/Notebook/August/12
From 2013.igem.org
(Difference between revisions)
(→1 - Electrophoresis of PCR products : RBS-BphR2 Part I, BphR2 Part I, BphR2 Part II, RBS-FNR Part I, FNR Part I and FNR Part II) |
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* Well 5 : 5µL FNR Part II+1µl of 6X loading dye | * Well 5 : 5µL FNR Part II+1µl of 6X loading dye | ||
* Well 6 : 5µL RBS-FNR Part I+1µl of 6X loading dye | * Well 6 : 5µL RBS-FNR Part I+1µl of 6X loading dye | ||
| - | * Well | + | * Well 7 : 5µL BphR2 Part I+1µl of 6X loading dye |
* Gel : 0.8% | * Gel : 0.8% | ||
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Revision as of 22:49, 1 October 2013
Notebook : August 12
Lab work
A - Aerobic/Anaerobic regulation system
Objective : characterize BBa_K1155000
1 - Digestion of BBa_K1155000 by SpeI/PstI, BBa_K1155007 and BBa_K1155003 by XBaI/PstI
Anaïs, Nadia, XiaoJing
Used quantities :
- BBa_K1155000 :
- Buffer FD : 5µL
- H2O : 38µL
- DNA : 5µL
- SpeI FD : 1µL
- PstI FD : 1µL
- BBa_K1155007 :
- Buffer FD : 5µL
- H2O : 23µL
- DNA : 20µL
- XBal FD : 1µL
- PstI FD : 1µL
- BBa_K1155003 :
- Buffer FD : 5µL
- H2O : 33µL
- DNA : 10µL
- XBal FD : 1µL
- PstI FD : 1µL
We let the digestion at 37°C during 15 minutes.
2 - Electrophoresis to check the digestion of BBa_K1155000 by SpeI/PstI, BBa_K1155007 and BBa_K1155003 by XBalI/PstI
Nadia
| File:Psdigestion12.jpg|350px]] |
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Expected sizes :
- Pfnr : ...
- RBS_LacZ_Term : 3500 kb
- RBS_AmilCP_Term : ...
- PSB1C3 : ...
|
We can't see any band for BBa_K1155000 digestion. The digestion failed. We will do it again. We obtain RBS-LacZ-Term and RBS-AmilCP-Term fragments. The digestion was good. We will purify it. |
3 - Digestion of BBa_K1155000 by SpeI/PstI
Anaïs, Nadia
Used quantities :
- Buffer FD : 5µL
- H2O : 38µL
- DNA : 5µL
- SpeI FD : 1µL
- PstI FD : 1µL
We let the digestion at 37°C during 15 minutes.
A - Aerobic/Anaerobic regulation system / B - PCB sensor system
Objective : obtaining FNR and BphR2 proteins
1 - Electrophoresis of PCR products : RBS-BphR2 Part I, BphR2 Part I, BphR2 Part II, RBS-FNR Part I, FNR Part I and FNR Part II
Damir
| [[]] |
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Expected size
- RBS-BphR2 Part I : 197 kb
- BphR2 Part II : 790 kb
- FNR Part I : 597 kb
- FNR Part II : 200 kb
- RBS-FNR Part I : 615 kb
- BphR2 Part I : 178 kb
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We can't see FNR Part I, FNR Part II and BphR2 Part i fragments at the good size. We will make the PCR again. We obtain RBS-BphR2 Part I, BphR2 Part II, RBS-FNR Part I frangments at the right size thanks to the PCR. We will purify it. |
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