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Team:Paris Saclay/Notebook/July/8
From 2013.igem.org
Contents |
Notebook : July 8
Summary:
FNR regulator system:
- performed another digestion for AmilCP, Pfnr plus sequence LacZ with Not I, Xho I, EcoR I and PST I.
- electrophoresis for those digest products
- enrichment culture for clone of the briobrick BBa_K1155000
- electrophoresis for extracted products of last friday
Lab work
Overnight incubation at 37°C with agitation.
The electrophoresis for the plasmids which we had extracted last week plus 3 other BioBrick promoter Bph R1, promoter Bph A1, Bph R2.
Volume added:
- Pfnr2: 10µl
- R1P: 10µl
- R2: 10µl
- A1P: 10µl
Migration at 100V for 25mins then continued till 45 minutes:
Results:
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The concentration of Pfnr2 was detected by NanoDrop :
- C = 1788.8 ng/µl
- 260/280 = 2.08
We considered that our extraction of plasmid DNA was not successful, we need to perform another extraction DNA.
Electrophoresis band size estimation
We used Clonemanager for band size estimation:
| Molecule | enzymes(Buffer) | Size |
| BBa_K1155000 | XHOI(Red) | 1289+892bp |
| BBa_K1155000 | ECoR I(Orange) | 2151bp |
Notebook : August 23
Lab work
A - Aerobic/Anaerobic regulation system
Objective : Bba_K1155003, Bba_K1155007
1 - Digestion of Bba_K592009, Bba_K1155007 by NotI, XhoI, EcoRI, EcoRI/PstI
Abou, Anaïs, Sheng, Zhou
Used quantities :
- NotI :
- Bba_K592009 : 2µL
- Buffer orange : 2µL
- NotI : 0.5µL
- H2O : 15.5µL
- XhoI :
- Bba_K592009 : 2µL
- Buffer red : 2µL
- XhoI : 0.5µL
- H2O : 15.5µL
- EcoRI :
- Bba_K592009 : 2µL
- Buffer orange : 2µL
- EcoRI : 0.5µL
- H2O : 15.5µL
- EcoRI/PstI :
- Bba_I732017 : 2µL
- Buffer orange : 2µL
- EcoRI : 0.5µL
- PstI : 0.5µL
- H2O : 15µL
We let our digestion 1h30 at 37°C.
2 - Electrophoresis of the digestion of Bba_K592009, Bba_I732017 by NotI, XhoI, EcoRI, EcoRI/PstI
Abdou, Anaïs, Sheng, Zhou
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Expected sizes :
- Bba_K592009 digested by NotI : 2046bp + 693bp
- Bba_K592009 digested by XhoI : 1842bp + 892bp
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