Patent ID: 6218152
Filing Date: 2001-04-17
Classification: C12N,C12Q

Abstract:
An in vitro method for amplifying a target polynucleotide region of an initial linear nucleic acid molecule, which method comprises the steps of:(A) forming, from said initial linear nucleic acid molecule, via enzymatic means, a partially single-stranded, partially double-stranded nucleic acid molecule, wherein one strand of said molecule contains a polynucleotide region that is complementary in sequence to said target polynucleotide region and is a circular strand lacking termini and the other of said strands is a non-circular strand having a 3' and a 5' terminus; said termini being hybridized to said circular strand of said molecule; said enzymatic means being an enzymatic reaction selected from the group consisting of a polymerase-mediated primer extension reaction, a recombination reaction and a ligation reaction, and(B) incubating said partially single-stranded, partially double-stranded nucleic acid molecule in the presence of a polymerase and under conditions sufficient to permit said polymerase to extend said 3' terminus of said non-circular strand in a template-dependent manner using said circular strand of said molecule as said template to thereby produce an extension product; wherein said template-dependent extension of said 3' terminus of said non-circular strand causes the strand displacement of said 5' terminus of said non-circular strand and results in said amplification of said target polynucleotide region.