Massive parallel method for decoding DNA and RNA

Patent metadata

Field	Value
Publication number	US-10669582-B2
Application number	US-201815915983-A
Country	US
Kind code	B2
Filing date	Mar 8, 2018
Priority date	Oct 6, 2000
Publication date	Jun 2, 2020
Grant date	Jun 2, 2020

Abstract

This invention provides methods for attaching a nucleic acid to a solid surface and for sequencing nucleic acid by detecting the identity of each nucleotide analogue after the nucleotide analogue is incorporated into a growing strand of DNA in a polymerase reaction. The invention also provides nucleotide analogues which comprise unique labels attached to the nucleotide analogue through a cleavable linker, and a cleavable chemical group to cap the —OH group at the 3′-position of the deoxyribose.

First claim

What is claimed is:

1. A deoxyribonucleic acid having attached by a phosphodiester bond at its 3′ end, a deoxyribonucleotide analogue having any one of the following structures:

wherein the O α is the 0 atom at the 3′ end of the deoxyribonucleic acid, and the wavy line represents the remainder of the deoxyribonucleic acid that is 5′ relative to the O α atom;
wherein R (a) represents a small, chemically cleavable, chemical group capping the oxygen at the 3′ position of the deoxyribose of the deoxyribonucleotide analogue, and (b) does not contain a ketone group;
wherein OR is not a methoxy group, an ester group or an allyl ether group;
wherein Tag represents a detectable fluorescent moiety;
wherein Y represents a chemically cleavable, chemical linker;
wherein the deoxyribonucleic acid:
i) produces a 3′—OH group on the deoxyribose of the deoxyribonucleotide analogue upon cleavage of R, and
ii) no longer includes a tag on the base upon cleavage of Y and wherein if the deoxyribonucleotide analogue is: (A), it is capable of forming hydrogen bonds with cytosine or a cytosine nucleotide analogue; (B), it is capable of forming hydrogen bonds with thymine or a thymine nucleotide analogue; (C), it is capable of forming hydrogen bonds with guanine or a guanine nucleotide analogue; or (D), it is capable of forming hydrogen bonds with adenine or an adenine nucleotide analogue.

2. The deoxyribonucleic acid of claim 1 , wherein the allyl ether group is O—CH 2 CH═CH 2 .

Assignees

• Univ Columbia

Inventors

• Ju Jingyue
• Li Zengmin
• Edwards John Robert
• Itagaki Yasuhiro

Classifications

• C12Q1/6874

  involving nucleic acid arrays, e.g. sequencing by hybridisation · CPC title

• C12Q2525/186

  incorporating a non-extendable or blocking moiety · CPC title

• C07H19/14

  Pyrrolo-pyrimidine radicals · CPC title

• C12Q1/686

  Polymerase chain reaction [PCR] · CPC title

• C12Q2535/101

  Sanger sequencing method, i.e. oligonucleotide sequencing using primer elongation and dideoxynucleotides as chain terminators · CPC title