Amplification of nucleic acids

摘要

The present invention relates to a method of amplifying a nucleic acid sequence present in a first strand of a double stranded nucleic acid molecule wherein said molecule incorporates an unmodified recognition site for a restriction enzyme capable of cutting the first strand at the 5' end of the sequence therein to be amplified and leaving the 3'-cut region of the second strand projecting beyond the cut site in the first strand. The method further comprises treating said molecule with said enzyme in the presence of a strand displacing polymerase and unmodified nucleotides for incorporation in an extending nucleic acid strand such that there is or becomes hybridised to said 3'-end region of the second strand a primer sequence complementary thereto whereby said primer sequence is extended in the 5' to 3' direction using the second strand as a template to regenerate the restriction endonuclease cut site and displace the sequence to be amplified.