Team:StanfordBrownSpelman/Lab Techniques11

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<h5><center>Steps Required</h5>
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<h5><center><a href="http://openwetware.org/wiki/Richard_Lab:Site_Directed_Mutagenesis" target="_blank">How to Make Mutagenesis Primers</a></h5>
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<b><a href="http://openwetware.org/wiki/Richard_Lab:Site_Directed_Mutagenesis" target="_blank">How to Make Mutagenesis Primers</a></b>
 
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<b>Double primer method:</b>
<b>Double primer method:</b>
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Latest revision as of 04:41, 16 October 2014

Stanford–Brown–Spelman iGEM 2014 — Amberless Hell Cell

How to Make Mutagenesis Primers
Double primer method:
Design mutagenesis primers. The targeted mutation should be included into both primers. The mutation can be as close as 4 bases from the 5-terminus. The mutation should be at least 8 bases from the 3-terminus. At least eight non-overlapping bases should be introduced at the 3-end of each primer. At least one G or C should be at the end of each primer. Design your primers (including the mutations) to have a Tm >=78°C.

Single primer method:
Design mutagenesis primer(s). The targeted mutation should be in the middle of the primer. Design your primers (including the mutations) to have a Tm >=78°C.

How to use thermal cycling:
Specifics can be found in the QuikChange Lightning Site-Directed Mutagenesis Kit.
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