Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extension.

@article{Horton1989EngineeringHG,
  title={Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extension.},
  author={Robert M. Horton and Horst Dieter MD. Hunt and S N Ho and Jeffrey K. Pullen and Larry R. Pease},
  journal={Gene},
  year={1989},
  volume={77 1},
  pages={61-8}
}
Gene splicing by overlap extension is a new approach for recombining DNA molecules at precise junctions irrespective of nucleotide sequences at the recombination site and without the use of restriction endonucleases or ligase. Fragments from the genes that are to be recombined are generated in separate polymerase chain reactions (PCRs). The primers are designed so that the ends of the products contain complementary sequences. When these PCR products are mixed, denatured, and reannealed, the… CONTINUE READING
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