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Team:Paris Liliane Bettencourt/Notebook/2011/09/13/
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===S38 in S24=== | ===S38 in S24=== | ||
Transformation failed. Positive control and negative control were good but no colonies for neither 20µL nor 200µL. | Transformation failed. Positive control and negative control were good but no colonies for neither 20µL nor 200µL. | ||
| + | ===Sequencing results=== | ||
| + | *S98 : clone 1 and 6 | ||
| + | *S99 : clone 2 | ||
| + | *S92 : the results are correct | ||
| + | ===Ligation & Transformation=== | ||
| + | *S79 in S27 | ||
| + | *S105 in S27 x2 (one of the MPs didn't show satisfactory sequencing results) | ||
| + | *S27 (-)ctrl for the two tubes that I used. (one concentrated at 9.8, the other at 5.4) | ||
| + | ===Minipreps=== | ||
| + | S105 x4 | ||
| + | S38 and pHM3 cultures launched for tomorrow. | ||
| + | ===Digestion & Gel extraction=== | ||
| + | cf. Cyrille | ||
| + | |||
| + | == Cyrille and Antoine == | ||
| + | |||
| + | Preparation of the last step of the ligation. | ||
| + | |||
| + | 12 miniprep of pHM3 were done and concentration measured. | ||
| + | 2 times 500 ng of each plasmid was digested in EP and runned on a gel. The bands where unclear. Only one miniprep seems to hold the correct insert altough there is a blur band for each of them at thee good size.this band was extracted and DNA was purified. | ||
| + | |||
| + | Treatment of the sequences - kinA project abandonned due to a wrongregsitry part | ||
| + | |||
| + | Digestion of PVeg ComS GA/PCR in EX, all the trnas (the digestion failed once), RBS frp ter, pT7 GFP in EX and SP (failed), S24 CI in EX, pVeg T7amber in EX (failed) and SP | ||
| + | |||
| + | The bands wer extracted and dna purified for some of them. The rest was freezed. | ||
| + | |||
| + | PHM3 was spread again on a plate to get single cones for minipreping | ||
| + | |||
| + | The two S24 comS from the transformation were insulated from the plate and grown for glycerol and MP. | ||
Latest revision as of 20:23, 13 September 2011
Contents |
Danyel
S38 in S24
Transformation failed. Positive control and negative control were good but no colonies for neither 20µL nor 200µL.
Sequencing results
- S98 : clone 1 and 6
- S99 : clone 2
- S92 : the results are correct
Ligation & Transformation
- S79 in S27
- S105 in S27 x2 (one of the MPs didn't show satisfactory sequencing results)
- S27 (-)ctrl for the two tubes that I used. (one concentrated at 9.8, the other at 5.4)
Minipreps
S105 x4 S38 and pHM3 cultures launched for tomorrow.
Digestion & Gel extraction
cf. Cyrille
Cyrille and Antoine
Preparation of the last step of the ligation.
12 miniprep of pHM3 were done and concentration measured. 2 times 500 ng of each plasmid was digested in EP and runned on a gel. The bands where unclear. Only one miniprep seems to hold the correct insert altough there is a blur band for each of them at thee good size.this band was extracted and DNA was purified.
Treatment of the sequences - kinA project abandonned due to a wrongregsitry part
Digestion of PVeg ComS GA/PCR in EX, all the trnas (the digestion failed once), RBS frp ter, pT7 GFP in EX and SP (failed), S24 CI in EX, pVeg T7amber in EX (failed) and SP
The bands wer extracted and dna purified for some of them. The rest was freezed.
PHM3 was spread again on a plate to get single cones for minipreping
The two S24 comS from the transformation were insulated from the plate and grown for glycerol and MP.
