Patent ID: 9605311
Date: 2017-03-28
CPC Classifications: C12Q

Claim:
1. A method of determining the sequence of a plurality of nucleic acid pairs, said method comprising: a) providing a sample comprising a plurality of double-stranded template nucleic acids having a template strand and a complementary strand; b) providing a solid support having a plurality of sites configured for single-molecule detection; c) for each of a plurality of template nucleic acids, linking the template strand and complementary strand by ligating a looped oligonucleotide to the double-stranded template nucleic acid to form a plurality of nucleic acid pairs comprising a looped structure that forms a physical tether between the template strand and the complementary strand, each nucleic acid pair comprising a free 3′ end and a free 5′ end; d) coupling the different nucleic acid pairs to the sites of the solid support, thereby spatially resolving the different pairs from each other; and e) determining the nucleotide sequence of the nucleic acid pairs on the solid support, wherein single molecules are detected individually, and wherein the template strand and the complementary strand remain physically tethered by the looped structure.