Team:Warsaw/Calendar-Stage1/14 September 2010
From 2010.igem.org
(Difference between revisions)
Line 10: | Line 10: | ||
<html> | <html> | ||
<!-- do not edit above me! --> | <!-- do not edit above me! --> | ||
+ | <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
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