Bi-directional sequencing compositions and methods

摘要

In some embodiments, methods for obtaining sequence information from a nucleic acid template linked to a support include hybridizing a first primer to a template strand linked to a support, sequencing a portion of the nucleic acid template, thereby forming an extended first primer product that is complementary to a portion of the nucleic acid template, In some embodiments, the method further includes introducing a nick into a portion of the template strand that is hybridized to the extended first primer product, degrading a portion of the template strand from the nick using a degrading agent, where a portion of the extended first primer remains hybridized to an undegraded portion of the template strand, and sequencing at least some of the single-stranded portion of the extended first primer by synthesis.

主权项

1. A method for generating single-stranded nucleic acids, comprising:
a) hybridizing a first primer to a nucleic acid molecule; b) extending the first primer via polymerization under polymerization conditions, thereby forming an extended first primer product that is complementary to a portion of the nucleic acid molecule; c) introducing a nick into a portion of the nucleic acid molecule that is hybridized to the extended first primer product, where the nick includes a free 5′ end and a free 3′ end in the nucleic acid molecule; and d) degrading the nucleic acid molecule from the free 5′ end of the nick using a degrading agent, thereby generating a single-stranded extended primer product which remains hybridized to an undegraded portion of the nucleic acid molecule.