Compositions and techniques for nucleic acid primer extension

By forming a stable ternary complex through specific contact between the primer-template nucleic acid hybrid and the polymerase and nucleotide mixture, the next base can be detected and distinguished from other base types in the template. This solves the problem of insufficient speed and accuracy of nucleic acid sequencing in existing technologies, and realizes rapid and accurate nucleic acid sequence feature labeling and diagnosis.

CN112074603BActive Publication Date: 2026-03-24PACIFIC BIOSCIENCES OF CALIFORNIA INC
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Patent Information

Application Number
CN201980014200.3
Authority / Receiving Office
CN · China
Patent Type
Patents(China)
Current Assignee / Owner
Priority Date
2018-02-06
Filing Date
2019-02-01
Publication Date
2026-03-24
Estimated Expiration
2039-02-01

AI Technical Summary

Technical Problem

Existing nucleic acid sequencing methods are not fast or accurate enough in clinical applications, leading to diagnostic delays and affecting patients' health and treatment options.

Method used

A method for characterizing nucleic acids is employed, which involves contacting a primer-template nucleic acid hybrid with a mixture of polymerase and nucleotides to generate an extended primer hybrid, which is then contacted with the polymerase and nucleotide mixture to form a stable ternary complex. The method detects the next base and distinguishes it from other base types in the template, thus determining the presence of base multivariates.

Benefits of technology

It improves the accuracy and speed of nucleic acid sequencing, reduces exposure to light and chemicals, increases read length and throughput, provides rapid nucleic acid sequence characterization, and supports rapid diagnostic and treatment decisions.

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Abstract

A method of characterizing a nucleic acid, the method comprising the steps of: (a) contacting a primer-template nucleic acid hybrid with a polymerase and a mixture of nucleotides to produce an extended primer hybrid and form a stable ternary complex comprising the extended primer hybrid, the polymerase, and a nucleotide cognate of a next base in the template, wherein the mixture contains nucleotide cognates of four different base types, wherein the nucleotide cognate of a first base type has a reversible terminator, and wherein nucleotide cognates of second, third, and fourth base types are extendable; (b) detecting the stable ternary complex to distinguish the next base from other base types in the template; and (c) determining the presence of a base multiplet in the template nucleic acid, the base multiplet comprising the first base type followed by the next base.
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