CPC C12Q 1/6853 (2013.01) [C12Q 1/6869 (2013.01)] | 19 Claims |
1. A method of characterizing a nucleic acid, comprising
(a) extending a primer-template nucleic acid hybrid comprising a primer and a template nucleic acid with extendable nucleotide cognates and a nucleotide cognate comprising a reversible terminator moiety to produce an extended primer-template hybrid comprising the reversible terminator moiety;
(b) forming a stabilized ternary complex comprising the extended primer-template hybrid, a polymerase and a nucleotide cognate of the next base in the template nucleic acid;
(c) detecting the stabilized ternary complex to distinguish the next base;
(d) deblocking the extended primer-template hybrid;
(e) repeating steps (a) through (d) using the extended primer-template hybrid as the primer-template nucleic acid hybrid to determine a series of base multiplets in the template nucleic acid;
(f) repeating steps (a) through (e) to determine at least 3 different series of base multiplets in the template nucleic acid each time; and
(g) combining at least four different series of base multiplets to determine a sequence of the nucleic acid.
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