Patent ID: 8765381
Filing Date: 2014-07-01
Classification: C12Q

Abstract:
1. A method for sequencing of first and second regions of target polynucleotides having different sequences, the method comprising: (a) providing beads comprising immobilized first strands of the target polynucleotides; (b) hybridizing a sequencing primer in solution to the immobilized first strands and sequencing the first region of the first strands of the of the target polynucleotides, wherein the sequencing comprises extension of a sequencing primer by sequential additions of nucleotides to generate an extended sequencing primer; (c) copying at least a portion of the first strands of the target polynucleotides by further extending the extended sequencing primer, thereby generating second strands having the second regions of the target polynucleotides; (d) removing a portion of the first strands from the beads, thereby generating an immobilized extension primer having a free 3′-hydroxyl; and (e) sequencing the second region of the second strands of the target polynucleotides using the immobilized extension primer, wherein determining the sequence of the first and second regions of the target polynucleotides achieves sequencing of the first and second regions of the target polynucleotides.