Patent ID: 8673560
Filing Date: 2014-03-18
Classification: C12N,C12Q

Abstract:
1. A method for amplifying a plurality of different target sequences within a sample, comprising: a) producing at least one hundred different amplified target sequences by amplifying at least one hundred different target sequences within a single amplification reaction mixture, wherein the producing includes contacting the at least one hundred different target sequences with a plurality of target-specific primers and a polymerase under amplification conditions and forming a plurality of double stranded amplified target sequences containing a primer-derived sequence, and wherein at least one of the plurality of target-specific primers and at least one of the primer-derived sequences includes a cleavable group; b) cleaving the cleavable group of at least one primer-derived sequence of at least one double stranded amplified target sequence, thereby forming a cleaved amplified target sequence; c) producing one or more double-stranded adapter-ligated amplified target sequences by ligating at least one double-stranded adapter containing an adapter sequence to the cleaved double-stranded amplified target sequence in a blunt-ended ligation reaction; and d) reamplifying at least one of the double-stranded adapter-ligated amplified target sequences using primers that bind to at least some portion of the adapter sequence; wherein the method includes use of no more than two target-specific primers to produce any one of the double stranded adapter-ligated amplified target sequences.