IGEM:metu/2009/Notebook/wound dressing/2009/09/09

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 * style="background-color: #EEE"|[[Image:igem-logo-150px.png|150px]] WOUND DRESSING
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 * style="background-color: #F2F2F2" align="center"|  |Main project page


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09.09.2009
1. PCR was done. Preparation of master mix. 11 plasmids + 1 control group 10x tag buffer 30 microliter 2 mM dNTP      30 microliter 25 mM MgCl2    30 microliter 5 mM t-primer  30    " 5 mM r-primer   30    " 1 u/µl tag Enz 12    " 2,5 u/µl pfu    0.3   " water nuclease free --- 185.7 µl aliquot as 49 µl and 1 µl DNA was add.

preparation of dNTP dATP -- 10 µl dTTP -- 10 µl dGTP -- 10 µl dCTP -- 10 µl water -- 460 µl

1- initial denaturation - 95 degree centigrade -- 3 minutes 2- denaturation - 95 degree centigrade -- 30 seconds 3- Annealing - 54 degree centigrade -- 30 seconds 4- Extension ( elongation step) - 72 degree centigrade -- 45 seconds 5- Final elongation - 72 degree centigrade -- 10 minutes (to ensure that any remaining single-stranded DNA is fully extended)

2.PCR purification (clean-up) was done.

3. Electrophorese of PCR products