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Team:Paris Liliane Bettencourt/Notebook/2011/08/06/
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== Cyrille == | == Cyrille == | ||
The ligation has given 7 transformant. 2 on the 3-5 200µL plate, 4 on the 6789 200µL plate and one on the 10-11 200µL plate | The ligation has given 7 transformant. 2 on the 3-5 200µL plate, 4 on the 6789 200µL plate and one on the 10-11 200µL plate | ||
| - | 7 miniprep were prepared from these clones. | + | 7 miniprep were prepared from these clones with an overday growth. |
'''Digestion:''' | '''Digestion:''' | ||
| Line 10: | Line 12: | ||
The mix contain: | The mix contain: | ||
| - | * 1µL 10x NEBuffer | + | * 1µL 10x NEBuffer 3 |
* 2µL plasmid | * 2µL plasmid | ||
| - | * 1µL | + | * 1µL EcoRV |
* 1µL EcoRI | * 1µL EcoRI | ||
* 5µL H2O | * 5µL H2O | ||
| + | |||
| + | After digesting one hour, the result was loaded on a gel and a picture was taken. Since the results are difficult to interpret, they where treated the day after. | ||
| + | |||
| + | [[File:CP0608Gel1.jpg|thumb|center|gel one with few BET. Wells: ladder, -, 1, 2, 3, 4, 5, 6, 7, ladder]] | ||
| + | [[File:CP0608Gel2.jpg|thumb|center|gel one with more BET. Wells: ladder, -, 1, 2, 3, 4, 5, 6, 7, ladder]] | ||
| + | |||
| + | It seems that it has worked for the well 4. | ||
Latest revision as of 16:08, 27 August 2011
Cyrille
The ligation has given 7 transformant. 2 on the 3-5 200µL plate, 4 on the 6789 200µL plate and one on the 10-11 200µL plate
7 miniprep were prepared from these clones with an overday growth.
Digestion:
The obtained plasmids are digested by both EcoRI and MscI from NEB in 10x NEBuffer 4.
The mix contain:
- 1µL 10x NEBuffer 3
- 2µL plasmid
- 1µL EcoRV
- 1µL EcoRI
- 5µL H2O
After digesting one hour, the result was loaded on a gel and a picture was taken. Since the results are difficult to interpret, they where treated the day after.
It seems that it has worked for the well 4.
