Patent ID: 9476080
Filing Date: 2016-10-25
CPC Classification: C12P,C12Q

Claim Text:
1. A method for amplifying a plurality of nucleic acid templates, comprising: a) providing a plurality of forward primers immobilized on a support, wherein the plurality of forward primers includes a first forward primer and a second forward primer, wherein the plurality of forward primers share 75% sequence identity; b) providing a nucleic acid reverse strand from the plurality of nucleic acid templates, having a forward primer-binding sequence that can hybridize to any one of the plurality of forward primers; c) hybridizing the first forward primer to the forward primer-binding sequence on the nucleic acid reverse strand; d) generating an extended forward strand that is substantially complementary to the nucleic acid reverse strand and is hybridized thereto, by primer extension of the first forward primer using the reverse strand as a template; e) partially denaturing the first forward primer and the forward primer-binding sequence on the nucleic acid reverse strand and hybridizing the second forward primer to the forward primer-binding sequence on the nucleic acid reverse strand, wherein a portion of the nucleic acid reverse strand is also hybridized to the extended forward strand; f) generating another extended forward strand that is substantially complementary to the nucleic acid reverse strand and is hybridized thereto, by primer extension of the second forward primer using the reverse strand as a template; and g) amplifying the plurality of nucleic acid templates simultaneously in a single continuous liquid phase without first compartmentalizing, by performing one or more amplification cycles comprising steps (e)-(f) under isothermal conditions to form clonal nucleic acid populations, wherein the second forward primer of step (e) of an amplification cycle acts as the first forward primer of step (e) of a subsequent amplification cycle; and wherein the amplifying is carried out by adjusting the isothermal conditions to a temperature that is higher than the T