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Team:Paris Saclay/Notebook/July/8
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Overnight incubation at 37°C with agitation. | Overnight incubation at 37°C with agitation. | ||
| + | <br> | ||
The electrophoresis for the plasmids which we had extracted last week | The electrophoresis for the plasmids which we had extracted last week | ||
Volume added: | Volume added: | ||
| - | Pfnr2 R1P R2 | + | *Pfnr2: 10µl |
| - | 10µl | + | *R1P: 10µl |
| + | *R2: 10µl | ||
| + | *A1P: 10µl | ||
| + | |||
| - | + | Migration at 100V for 25mins then continued till 45 minutes: | |
| - | + | <br< | |
| + | Results: | ||
The concentration of Pfnr2 was detected by NanoDrop : | 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. | ||
Revision as of 09:56, 21 September 2013
Notebook : July 8
Summary:
FNR regulator system:
- performed another digestion for AmilCP 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
Restriction digest
For some of the enzymes which are possible to use one common buffer, we did double digest
| DNA | enzyme | buffer |
| AmilCP1 | Not I | Orange |
| AmilCP2 | Not I | Orange |
| AmilCP1 | Xho I | Red |
| AmilCP2 | Xho I | Red |
| AmilCP1 | EcoR I | Orange |
| AmilCP2 | EcoR I | Orange |
| LacZ1 | EcoR I + PST I | Orange |
| LacZ2 | EcoR I + PST I | Orange |
Volume added:
| DNA | 2(µl) |
| buffer | 2(µl) |
| Enzyme(foreach) | 0.5(µl) |
| H20 | 15.5(µl) |
| total | 20(µl) |
The electrophoresis (135 V for 30 minutes) for digest products were migrated in agarose gel (0.5X).
Results:
Overnight incubation at 37°C with agitation.
The electrophoresis for the plasmids which we had extracted last week
Volume added:
- Pfnr2: 10µl
- R1P: 10µl
- R2: 10µl
- A1P: 10µl
Migration at 100V for 25mins then continued till 45 minutes:
<br<
Results:
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.
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