Paris/July 31
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| + | [[Paris/July 30|yesterday]] -- [[Paris/August 1|tomorrow]] <br> | ||
| + | == pTOPO cloning == | ||
| + | Cloning of PCR products: P5, P6, P7, P8, P9, P10 (see [[Paris/Freezer]]) | ||
| + | |||
| + | |||
| + | == Transformation == | ||
| + | |||
| + | Ligations from yesterday | ||
| + | |||
| + | |||
| + | |||
== PCR mutagenesis DGAT == | == PCR mutagenesis DGAT == | ||
| Line 20: | Line 31: | ||
|Success= YES | |Success= YES | ||
|Image= DGAT107312007.jpg | |Image= DGAT107312007.jpg | ||
| - | |Band= | + | |Band= 3, 4, 5, 6 (11 = control without DNA matrix) |
|}} | |}} | ||
<br> | <br> | ||
| Line 41: | Line 52: | ||
|Success= YES | |Success= YES | ||
|Image= DGAT107312007.jpg | |Image= DGAT107312007.jpg | ||
| - | |Band= | + | |Band= 7,8,9,10 (11 = control without DNA matrix) |
|}} | |}} | ||
<br> | <br> | ||
Latest revision as of 17:52, 7 October 2007
pTOPO cloning
Cloning of PCR products: P5, P6, P7, P8, P9, P10 (see Paris/Freezer)
Transformation
Ligations from yesterday
PCR mutagenesis DGAT
DGAT has Pst1 restriction site. We want to mutagenize it to do a biobrick.
| PCR : DGAT1 | ||||||
|---|---|---|---|---|---|---|
| PCR Settings | Buffer (5x) | 5x 10µL | Expected size | |||
| Annealing (°C) | MgCl2 10µM | 10µM 0µL | 1087 | |||
| 50 | dNTP 10µM | 10µM 1µL | Success | |||
| Time Elongation | Oligo F 10µM | 27 ForDGAT1 | 2.5µL | YES | ||
| 2m00' | Oligo R 10µM | 13 DPst1-DGAT-R | 2.5µL | Image (click to enlarge) | ||
| Number cycles | Water | 33µl | 
| |||
| 35 | Polymerase | Phusion 0.5µL | Band (0=ladder) | |||
| DNA | Miniprep pKS::DGAT (1µL = 138ng) | 3, 4, 5, 6 (11 = control without DNA matrix) | ||||
| PCR : DGAT2 | ||||||
|---|---|---|---|---|---|---|
| PCR Settings | Buffer (5x) | 5x 10µL | Expected size | |||
| Annealing (°C) | MgCl2 10µM | 10µM 0µL | 396 | |||
| 50 | dNTP 10µM | 10µM 1µL | Success | |||
| Time Elongation | Oligo F 10µM | 12 DPst1-DGAT | 2.5µL | YES | ||
| 2m00' | Oligo R 10µM | 27 Rev-DGAT1 | 2.5µL | Image (click to enlarge) | ||
| Number cycles | Water | 33µl |
| |||
| 35 | Polymerase | Phusion 0.5µL | Band (0=ladder) | |||
| DNA | Miniprep pKS::DGAT (1µL = 138ng) | 7,8,9,10 (11 = control without DNA matrix) | ||||
