Team:Warsaw/Calendar-Stage1/14 September 2010

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<h2>Mutagenesis of MinC</h2>
<h4>Kuba</h4>
<h4>Kuba</h4>
<p align="justify">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.
<p align="justify">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.

Revision as of 14:51, 25 October 2010

Example Tabs

Mutagenesis of MinC

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