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US 2016/0237486 A1Published application
Native-extension parallel sequencing
AssigneeCentrillion Technology Holdings Corporation
InventorsWei Zhou, Rui Mei, Julian Lucas, Guochun Liao
Life of this patent
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Abstract
The present invention provides methods for native extension parallel sequencing of polynucleotide.
Claims15
A method of sequencing a target nucleic acid, comprising:
- (a) providing a plurality of substrates, wherein a target nucleic acid is immobilized to said plurality of substrates;
- (b) hybridizing said target nucleic acid on said plurality of substrates with a sequencing primer;
- (c) extending at least one of the said plurality of substrates with one, two or three different nucleotides and a polymerase;
- (d) sequencing one or more bases of the target nucleic acid from the primer or extended primer;
- and
- (e) assembling target nucleic acid sequence based upon said sequencing.
3 more elements
A method of sequencing a plurality of target nucleic acid molecules, comprising:
- (a) providing a plurality of substrates, wherein said target nucleic acids are immobilized to said plurality of substrates;
- (b) hybridizing said target nucleic acids on said plurality of substrates with a sequencing primer;
- (c) extending at least one of the said plurality of substrates with one, two or three different nucleotides and a polymerase;
- (d) sequencing one or more bases of the target nucleic acids from the primer or extended primer;
- and
- (e) assembling target nucleic acid sequences based upon said sequencing.
3 more elements
A method of sequencing a target nucleic acid molecule, comprising:
- (a) providing a plurality of substrates, wherein the total number of substrates is n, each substrate is designed as substrate (i) and i is an integer from 1 to n, wherein each of said substrates comprises a capture site comprising a capture probe, and wherein each of said capture probe comprises a sequence that is complementary to a target nucleic molecule;
- (b) forming a plurality of hybridization complexes on said substrate, each complex comprises:
- said capture probe and a copy of said target nucleic acid molecule;
- (c) extending said capture probes on each said substrate (i) by repeating i−1 times the step of:
- contacting said hybridization complex on said substrate (i) sequentially with dATP, dCTP, dGTP and dTTP in the presence of a polymerase, thereby extending said capture probes by one or more bases using said target nucleic acid molecules as templates;
- (d) contacting said each hybridization complex on each said substrate (i) sequentially with one of labeled dATP, dCTP, dGTP and dTTP in the presence of a polymerase;
- (e) detecting the incorporation of said of labeled dATP, dCTP, dGTP and dTTP to each of said capture probe on each of said substrate (i) to obtain a sequence read from each substrate (i);
- and
- (f) determining the sequence of said target nucleic acid molecule by assembling said sequence reads.
6 more elements
Dependencies are inferred from the claim text.