Patent ID: 8247177
Filing Date: 2012-08-21
Classification: C12Q

Abstract:
1. A method for pairwise sequencing of target double-stranded polynucleotides, the method comprising: (a) providing a surface comprising a plurality of target double-stranded polynucleotides comprising first and second template strands, the first and second template strands comprising first and second regions, respectively, to be sequenced; (b) forming a single-stranded region in the first template strands and hybridizing first sequencing primers to the single-stranded region in the first template strands; (c) carrying out a first sequencing reaction by sequential addition of nucleotides to the first sequencing primer to determine the sequence of the first region of the first template strands; (d) cleaving the first template strands; (e) removing all or a portion of the first template strands; (f) hybridizing a second sequencing primer to the second template strands; (g) carrying out a second sequencing reaction by sequential addition of nucleotides to the second sequencing primer to determine the sequence of the second region of the second template strands, wherein sequencing of the first and second regions permits pairwise sequencing of the target double-stranded polynucleotides.