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Team:Paris Liliane Bettencourt/Notebook/2011/09/03/
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==Danyel== | ==Danyel== | ||
*Miniprep of '''S92'''(Pveg_spoVG_tRNA_Ter in pHM3 in MH1 cells). | *Miniprep of '''S92'''(Pveg_spoVG_tRNA_Ter in pHM3 in MH1 cells). | ||
*Gel extraction of S82 digested on S & P and S82 digested on E & S | *Gel extraction of S82 digested on S & P and S82 digested on E & S | ||
*Ligation of: | *Ligation of: | ||
| - | **S38 in S24 ( | + | **S38 in S24 (AmpR) |
**S38 in S82 (CmR) | **S38 in S82 (CmR) | ||
**S58 in S82 (CmR) | **S58 in S82 (CmR) | ||
**(with negative controls for S24 and S82) | **(with negative controls for S24 and S82) | ||
at ratio 6:1 and 100:1 | at ratio 6:1 and 100:1 | ||
| + | All ligations were transformed then plated. | ||
| + | |||
| + | ===GFP amber=== | ||
| + | Transformation of the digested Quick change PCR products and plating. However, due to a stupid mistake that I'm a little to embarrassed to mention (didn't add LB before 1hour incubation), the transformation will probably have to be redone. | ||
| + | |||
| + | == Cyrille == | ||
| + | |||
| + | === Preparation of new competent cell Turbo === | ||
| + | |||
| + | === Digestion of pVeg YFP TetR and pHM3 === | ||
| + | |||
| + | Minipreped from the glycerol S90 in the morning | ||
| + | |||
| + | pVEg YFP TetR was digested in 20 µL in EP and SP | ||
| + | pHM3 was digested in EP. The gel gives strange results | ||
| + | |||
| + | [[File:CP0309_gel.jpg|center|thumb|YFP TetR SP and XP]] | ||
| + | [[File:CP0309_pHM3.jpg|center|thumb|pHM3.jpg]] | ||
| + | |||
| + | Gel runned and bands extracted. | ||
| + | |||
| + | [[File:CP0309_gel_cutted.jpg|center|thumb]] | ||
| + | |||
| + | === Glycerols of 4,8,2 and TetO in pHM3 === | ||
| + | |||
| + | Transformation of the miniprep was done and plated. | ||
| + | |||
| + | === Test of a for contamination in competent cells === | ||
| + | |||
| + | One tube of competent cells was growth for two hours in LB and plated on ampicilyn plates. This is a check for contaminations by ampicilyn resistant strains. | ||
Latest revision as of 22:39, 3 September 2011
Contents |
Danyel
- Miniprep of S92(Pveg_spoVG_tRNA_Ter in pHM3 in MH1 cells).
- Gel extraction of S82 digested on S & P and S82 digested on E & S
- Ligation of:
- S38 in S24 (AmpR)
- S38 in S82 (CmR)
- S58 in S82 (CmR)
- (with negative controls for S24 and S82)
at ratio 6:1 and 100:1 All ligations were transformed then plated.
GFP amber
Transformation of the digested Quick change PCR products and plating. However, due to a stupid mistake that I'm a little to embarrassed to mention (didn't add LB before 1hour incubation), the transformation will probably have to be redone.
Cyrille
Preparation of new competent cell Turbo
Digestion of pVeg YFP TetR and pHM3
Minipreped from the glycerol S90 in the morning
pVEg YFP TetR was digested in 20 µL in EP and SP pHM3 was digested in EP. The gel gives strange results
Gel runned and bands extracted.
Glycerols of 4,8,2 and TetO in pHM3
Transformation of the miniprep was done and plated.
Test of a for contamination in competent cells
One tube of competent cells was growth for two hours in LB and plated on ampicilyn plates. This is a check for contaminations by ampicilyn resistant strains.
