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EPF-Lausanne/24 September 2009
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==Wet Lab== | ==Wet Lab== | ||
===Transformation in Trp K.O.=== | ===Transformation in Trp K.O.=== | ||
| - | The 3 newly competent strains (JW4356-2, JRG 1046, JRG 465) will be transformed | + | The 3 newly competent strains (JW4356-2, JRG 1046, JRG 465) will be transformed with : |
| - | + | - RO2#4 + BB1 (+Amp, Kana) -> double transformation. | |
| + | - RO2#4 (+Kana) | ||
| + | |||
| + | - RO1#1 (+Kana) | ||
| + | |||
| + | - RO1#1 + BB1 (+Amp, Kana) -> double transformation. | ||
| + | |||
| + | Note : the 3 control (1 ug/ul DNA, 100 pg/ul DNA, 10 pg/ul DNA -> 10ul for each transf) + KANA. We will count the nb of clones the next morning on the plates to determine transformation efficiency. | ||
| + | |||
| + | The normal protocol for these transformation has been used (50 ul cells + 10 ul total volume DNA) : 20 min on ice, heat shoch (45 s at 42°C), SOB (500ul) at 37°C for 1 h, plated on according antibiotic LB/Agar plates. Put overnight at 37°C. | ||
| + | |||
| + | Note : the double transformation : the whole tue (about 560 ul) is added to the plates. The single transformation : only 150 ul of the mixture. | ||
| + | |||
| + | ==People in the lab== | ||
| + | Mél, Nicolas, Christian, Basile | ||
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Latest revision as of 15:21, 4 October 2009
Wet Lab
Transformation in Trp K.O.
The 3 newly competent strains (JW4356-2, JRG 1046, JRG 465) will be transformed with :
- RO2#4 + BB1 (+Amp, Kana) -> double transformation.
- RO2#4 (+Kana)
- RO1#1 (+Kana)
- RO1#1 + BB1 (+Amp, Kana) -> double transformation.
Note : the 3 control (1 ug/ul DNA, 100 pg/ul DNA, 10 pg/ul DNA -> 10ul for each transf) + KANA. We will count the nb of clones the next morning on the plates to determine transformation efficiency.
The normal protocol for these transformation has been used (50 ul cells + 10 ul total volume DNA) : 20 min on ice, heat shoch (45 s at 42°C), SOB (500ul) at 37°C for 1 h, plated on according antibiotic LB/Agar plates. Put overnight at 37°C.
Note : the double transformation : the whole tue (about 560 ul) is added to the plates. The single transformation : only 150 ul of the mixture.
People in the lab
Mél, Nicolas, Christian, Basile
