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Team:Paris Saclay/Notebook/July/8
From 2013.igem.org
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| - | + | {| align="center" | |
| - | + | | style="width:350px;border:1px solid black;" | [[File:PS080713dig.jpg|center|350px]] | |
| + | | style="width:350px;border:1px solid black;" | | ||
| + | *Well 1,12 : marker | ||
| + | *Well 2: AmilCP1 + Ecor I | ||
| + | *Well 3: AmilCP2 + Ecor I | ||
| + | *Well 4: AmilCp1 + Not I | ||
| + | *Well 5: AmilCP2 + Not I | ||
| + | *Well 6: AmilCp1 + Xho I | ||
| + | *Well 7: AmilCp1 + Xho I | ||
| + | *Well 8: LacZ 1 + Ecor I + Pst I | ||
| + | *Well 9: LacZ 2 + Ecor I + Pst I | ||
| + | *Well 10: Nothing | ||
| + | *Well 11: fnr promoter | ||
| + | *gel 0.8% | ||
| + | |}<br> | ||
Revision as of 10:04, 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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