Method For Gene Splicing By Using Overlap Extension (SOE)

Reference #:

1988-032

Inventors/Contributors

Steffan Ho, M.D., Ph.D., Robert Horton, Ph.D.

Description

A method is provided for joining two DNA molecules by first amplifying them by means of polymerase chain reactions (PCR) carried out on each molecule using oligonucleotide primers designed so that the ends of the resultant PCR products contain complementary sequences. When the two PCR products are mixed, denatured and reannealed, the single-stranded DNA strands having the complementary sequences anneal and then act as primers for each other. Extension of the annealed area by DNA polymerase produces a double-stranded DNA molecule in which the original molecules are spliced together. Gene splicing by overlap extension (SOE), provides for recombining DNA molecules at precise junctions irrespective of nucleotide sequences at the recombination site and without the use of restriction endonucleases or ligase. The SOE approach is a fast, simple, and extremely powerful, way of recombining and modifying nucleotide sequences.

Patent Status

	U.S. Patent Number 5,023,171 Issued June 11, 1991