METHOD OF GENERATING LONG NUCLEIC ACID MOLECULES OF DEFINED SEQUENCE

摘要

A method of generating a single-stranded nucleic acid of a predetermined sequence, comprising: a) contacting a poynucleotide comprising a 5' portion and a 3' portion, a template oligonucleotide having a 3' blocker, a 5' template portion, and a 3' portion which is sufficiently complementary to the 3' portion of the polynucleotide to hybridize thereto, and a polymerase under conditions in which the 3' portion of the polynucleotide hybridizes to the 3' portion of the template oligonucleotide and elongates to produce an elongated polynucleotide product; b) denaturing the duplex comprising the elongation product and a template oligonucleotide; c) repeating a) and b) in the presence of a second or subsequent template oligonucleotide to produce a second or subsequent elongation polynucleotide product.