User:Maximilian Peters/site-directed-mutagenesis

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(New page: ==Site-directed mutagenesis protocol== This protocol does not require any special enzymes or kits, only a regular PCR block and equipment to handle DNA. 1. Primer design with Strategene'...)
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Revision as of 11:05, 8 June 2011

Site-directed mutagenesis protocol

This protocol does not require any special enzymes or kits, only a regular PCR block and equipment to handle DNA.

1. Primer design with Strategene's primer design tool 2. Order both primers 5'-phosphorylated 3. Perform a 20 microL PCR reaction to test the primers, if a strong band in the desired region is obtained repeat the PCR with 2-4 50x microL reactions 4. Run the PCR product on an Agarose gel (1% or less) and isolate the desired band with a method of your choice 5.

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