Abstract: The present invention concerns methods of assembly of a polynucleotide on a solid-support by performing steps of annealing, ligation, and extension. The steps may be repeated in a cyclical manner to assemble immobilized double- or single-stranded polynucleotides with functional gene properties. The immobilized polynucleotides may be amplified by the polymerase chain reaction, and detected and quantitated by an exonuclease assay with a self-quenching, fluorescent probe. The polynucleotide may be cleaved from the solid-support by chemical or enzymatic cleavage.
Type:
Grant
Filed:
November 12, 1998
Date of Patent:
August 24, 1999
Assignee:
The Porkin-Elmer Corporation
Inventors:
Michael W. Hunkapiller, Andrew C. Hiatt