Recombinant circle PCR and recombination PCR for site-specific mutagenesis without PCR product purification.

@article{Jones1992RecombinantCP,
  title={Recombinant circle PCR and recombination PCR for site-specific mutagenesis without PCR product purification.},
  author={D. Holstead Jones and Stanley C. Winistorfer},
  journal={BioTechniques},
  year={1992},
  volume={12 4},
  pages={528-30, 532, 534-5}
}
Two simple methods for site-specific mutagenesis are described and compared. In each method, the PCR is used in two separate amplifications to mutate the site of interest and to add ends to one PCR product that are homologous to the ends of the other PCR product. In the first method, the two products are combined, denatured and reannealed prior to transformation of E. coli in order to form recombinant circles in vitro, while in the second method, the two linear products are co-transfected… CONTINUE READING