- 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.