Team:Warsaw/Calendar-Stage1/14 September 2010
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+ | <p align="justify">PCR product was purified by agarose electrophoresis and digested over-night with DpnI | ||
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Revision as of 14:46, 25 October 2010
Kuba
Directed mutagenesis was preformed on pSB-MinC in order to remove an additional PstI site. Two primers were used: MinCmutF (TAATTGCAGTCGCGCCGC) and MinCmutR (GTCGAAAACGCTTTGACCG). PCR was run with Yellow Pfu Polimerase (Eurex) in a standard reaction mix.
PCR conditions were as follows:
- initial denaturation 95 oC; 3'
- denaturation 95 oC; 30 s
- annealing 51 oC; 30 s
- elongation 72 oC<; 7'/li>
- x 30 cycles
- 72 oC; 10'
PCR product was purified by agarose electrophoresis and digested over-night with DpnI