Team:University of Sheffield/Gosia

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16/08/08

Primers for Kan cassette synthesis ordered.

21/08/08

Primers arrive. Rehydration of a primers was carried as follows: volume a dH2O (uL) equal to ten times the number of nanomoles of DNA present in the tube will produce a stock solution at 100µM concentration.

Due to data corruption I do not have an electronic copy of my lab book any more. I am publishing my handwritten notes as an evidence of my work.

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gp1
gp2
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24/10/08

BBs construction continues - a PCR was run to PCR-out genes cqsS, barA and Red Recombinase cassette. A PCR run yesterday by Rosie didn't work in all cases (Illustration 1).

The procedure was carried out in accordance with Protocol [X] except that 1 µl of 100 mM stock solutions of forward and reverse primers were used instead of 2 pm/µl as usually. 10 ng of plasmid pKD46 was added instead of 1 ng. Exact volumes of PCR reaction mixture components is listed in Table 1.

[[Image:tabe1.jpg|center|tabtab]

PCR of BBs carried out by Rosie yesterday (23/10/08)

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