Endy:Site-directed Mutagenesis: Difference between revisions
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Revision as of 13:20, 1 July 2005
Primer Design
- See this page for general advice on primer design.
- Primers should contain the mutations to be introduced but should anneal to the sequence to be mutated. You will need a forward and reverse primer.
- Primers should be between 25 and 45 base-pairs in length.
- The mutation(s) should be in the middle of the primers.
- Mutation efficiency is greatly influenced by the purity of the primers. Its worth getting purified primers. No need for 5' phosphorylation.
- See the Stratagene manual for more detailed information.
Experimental Protocol
- See the Stratagene manual for the suggested protocols.
- Comments and usage notes on this protocol will be added when the protocol has been tested.