Team:Paris Liliane Bettencourt/Notebook/2011/09/06/

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Team IGEM Paris 2011

Contents

Cyrille

Transformations check

pVeg-YFP in pHM3: Transformation works. About the same ratio in 1:1 and 1:10. A bit less in 1:100. 

                  -> Transformation seems clean, control weak but correct
                       -> 10 colonies insulated

TetO-pVeg-YFP in pHM3: Transformation works, a bit less eficiency. It seems clean. (no negative control) 

                  -> A bit weaker, control correct
                       -> 10 colonies insulated

YFP TetR BB in pHM3: Negative control very polluted. About 1.3 times the number of colonies in the T1 and very few in the T2. Positive control correct. 

                       -> 10 colonies insulated from plate T1

ComS in pSB1C3: Negative control very clean (~20 colonies in the 200µL plate) Very few colonies on the other plates. Seems to have worked best with ratio 1:10-5. Very fw colonies on the other plates 

                      -> 10 colonies insulated


Test of new comptetent cells

Negative control on Ampicylin are clean Negative control (400µL out of 1ml plated)on Cm are a bit polluted. About 30 clones.

Positive control with tube one and two are very good, but they are not red!!! How come?

To conclude, use the MH1 cells (cap green) for cloning on Cm and Turbo cells to clone on Ampicilyn!!!

PCR colony

The clones insulated was also PCR colonysed. We used a Taq MasterMix.

An error was made while programming the PCR machine, and the PCR on pHM3 stayed for two hours at 95°C. THe PCR was atempter whatsoever.

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