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Team:Paris Saclay/Notebook/July/4
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* Well 1 to 4 : 5µL of PCR products with VF/VR primers+1µL of 6X loading dye | * Well 1 to 4 : 5µL of PCR products with VF/VR primers+1µL of 6X loading dye | ||
| - | * Well 5 to 6 : 5µL of PCR product withVF/Pfnr_Down primers+ | + | * Well 5 to 6 : 5µL of PCR product withVF/Pfnr_Down primers+1µL of 6X loading dye |
* Well 7 : 6µL of DNA Ladder | * Well 7 : 6µL of DNA Ladder | ||
| - | * Well 8 to 9 : 5µL of PCR product withVF/Pfnr_Down primers+ | + | * Well 8 to 9 : 5µL of PCR product withVF/Pfnr_Down primers+1µL of 6X loading dye |
* Well 10 to 13 : 5µL of PCR products with Pfnr_Up/VR primers+1µL of 6X loading dye | * Well 10 to 13 : 5µL of PCR products with Pfnr_Up/VR primers+1µL of 6X loading dye | ||
* Well 14 : 6µL of DNA Ladder | * Well 14 : 6µL of DNA Ladder | ||
Revision as of 17:36, 26 September 2013
Notebook : July 4
Lab work
A - Aerobic/Anaerobic regulation system
Objective : obtaining Bba_K1155000
1 - Estimation of Pfnr Colony PCR size fragments
Abdou, Anaïs, Sheng
We used software gene manager to find the correct size of our fragments.
Estimated size :
- VF/VR primer -> Plasmid without Pfnr size : 277bp
- VF/Pfnr_down -> Plasmid with Pfnr size : 276bp
- Pfnr_Up/VR -> Plasmid with Pfnr size : 311bp
2 - Electrophoresis of Colony PCR products
Zhou
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We obtain fragment at the right size in wells : 3, 4 , 8 , 9 12 and 13. We will digest our biobrick to confirm that Pfnr insert PSB1C3 correctly. |
3 - Stock of Bba_K1155000
Zhou
Used quantities :
- Bba_K1155000 confirmed : 1 mL
- Glycerol : 500µL glycerol.
We stocked them at -20°C.
4 - Extraction of Bba_K1155000 from DH5α
Anaïs, Sheng
Protocol : Hight copy plamid extraction
5 - Digestion of Bba_K1155000 by NotI, MluI, HpaI to check good insertion of Pfnr in PSB1C3
Abdou
Used quantities :
- Bba_K1155000 : 2µL
- Bufer oranger : 2µL
- NotI or MluI or HpaI : 0.5µL
- H2O : 15.5µL
We let the digestion 1h30 at 37°C.
6 - Culture of Bba_K1155000
Anaïs, Sheng
We made 2 cultures of bacterias transformed with Bba_K1155000 that show a fragments at the right size at electrophoresis.
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