Endy:Site-directed Mutagenesis

Primer Design

• See this page for general advice and software suggestions for 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.