Endy:Site-directed Mutagenesis
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==Primer Design== | ==Primer Design== | ||
| - | *See [[Designing Primers | this page]] for general advice | + | *See [[Designing Primers | 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 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. | *Primers should be between 25 and 45 base-pairs in length. | ||
Revision as of 11:09, 2 July 2005
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.
