Uppsala-Sweden/11 August 2009

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Evaluation of competent cells

Due to the run of bad transformations we've experienced we decided to evaluate the quality of our cells. We used our previosly extraxted Pbs1A (761 µg/ml) plasmid and as a control the pUC1G (100 pg/µl) plasmid from Invitrogen. The evaluation part of the protocol TOP10 chemically competent cells was followed with the deviations noted:

For transformation we used 5 µl of 100 pg/µl plasmid instead of the 1 µl of 10 pg/µl as told by the protocol, the Pbs1A was diluted down to this concentration. This as this was what was given in the protocol supplied by Invitrogen.

20 and 200 µl of the transfomed cells were plated on 50 µg/ml ampicillin plates and icubated at 37 degrees over nigth.

--Anders 15:35, 11 August 2009 (UTC)

Colony PCR

Colony PCR of new transformants was performed with Taq-Polymerase. Protocol will follow soon.

Inoculation of pSB1A3, pSB1AK3 and pSB1AC3

Cultures carrying pSB1A3, pSB1AK3 and pSB1AC3 were inoculated for Miniprep purposes at 37°C and 250rpm on LB.

--Karl.brune 17:06, 11 August 2009 (UTC)

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