EPF-Lausanne/24 September 2009

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==Wet Lab==
==Wet Lab==
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===Transformation in Trp K.O.===
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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)
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==People in the lab==
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- 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==
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Mél, Nicolas, Christian, Basile
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Latest revision as of 15:21, 4 October 2009



Contents

24 September 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



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