Patent ID: 6423495
Filing Date: 2002-07-23
Classification: C12Q

Abstract:
A method of amplifying a nucleic acid sequence present in a first strand of a double stranded nucleic acid molecule comprised of complementary first and second strands whereinsaid molecule incorporates an unmodified recognition site for a restriction enzyme capable of cutting the first strand at the 5â€² end of the sequence therein to be amplified and leaving the 3â€²-end region of the second strand projecting beyond the cut site in the first strand, and said method comprises treating said nucleic acid molecule with said enzyme in the presence of a strand displacing polymerase and unmodified nucleotides for incorporation in an extending nucleic acid strand under conditions such that there is or becomes hybridised to said 3â€²-end region of the second strand a primer sequence complementary thereto whereby said primer sequence is extended in the 5â€² to 3â€² direction using the second strand as a template to re-generate the restriction endonuclease cut site and displace the sequence to be amplified.