https://www.neb.com/applications/cloning-and-synthetic-biology/site-directed-mutagenesis

Alternatively, if you don't want to use the SDM mix: 

  • Design overlapping forward and reverse primers with the desired point mutation. 
  • PCR as normal (hybridization temperature should be lower than dimerization temperature/deltaG)
  • Transform PCR product (double-nicked plasmid)
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