Patent ID: 8124346
Filing Date: 2012-02-28
Classification: C12Q

Abstract:
1. A method for selectively amplifying a target nucleic acid sequence from a deoxyribonucleic acid or a mixture of nucleic acids, comprising the steps of: (a) performing a first-stage amplification of said target nucleic acid sequence at a first annealing temperature, said first-stage amplification comprising at least two cycles of primer annealing, primer extending and nucleic acid denaturing using a first primer pair, each primer of said first primer pair comprising a first 3′-end portion and a first 5′-end portion, and each primer of said first primer pair comprising: whereby said regulator sequence restricts primer annealing to said first 3′-end portion, and whereby an amplification product of said target nucleotide sequence is generated; and (b) performing a second-stage amplification of said amplification product generated from step (a) at a second annealing temperature, said second-stage amplification comprising at least one cycle of primer annealing, primer extending and nucleic acid denaturing using a second primer pair, each primer of said second primer pair comprising a pre-selected arbitrary nucleotide sequence corresponding to said first 5′-end portions of each primer used in step (a), under conditions in which each primer of said second primer pair anneals to 3′- and 5′-ends of said amplification product, respectively, whereby said amplification product is re-amplified; wherein the first annealing temperature and the second annealing temperature are different.