"
Team:Paris Saclay/Notebook/July/8
From 2013.igem.org
(Difference between revisions)
(→Lab work) |
(→Summary:) |
||
| Line 6: | Line 6: | ||
==''Summary:''== | ==''Summary:''== | ||
FNR regulator system: | FNR regulator system: | ||
| - | *performed another digestion for AmilCP plus sequence LacZ with Not I, Xho I, EcoR I and PST I. | + | *performed another digestion for AmilCP, Pfnr plus sequence LacZ with Not I, Xho I, EcoR I and PST I. |
*electrophoresis for those digest products | *electrophoresis for those digest products | ||
*enrichment culture for clone of the briobrick BBa_K1155000 | *enrichment culture for clone of the briobrick BBa_K1155000 | ||
*electrophoresis for extracted products of last friday | *electrophoresis for extracted products of last friday | ||
| - | |||
=='''Lab work'''== | =='''Lab work'''== | ||
Revision as of 10:05, 21 September 2013
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
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.
| Previous week | Back to calendar | Next day |
