Massive parallel method for decoding dna and rna

Patent metadata

Field	Value
Publication number	US-2018201642-A1
Application number	US-201815915983-A
Country	US
Kind code	A1
Filing date	Mar 8, 2018
Priority date	Oct 6, 2000
Publication date	Jul 19, 2018
Grant date	—

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

1 - 60 . (canceled)

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

wherein the O α is the O 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 or an ester 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 analog 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.

Assignees

• Univ Columbia

Inventors

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

Classifications

• C12Q1/686

  Polymerase chain reaction [PCR] · CPC title

• C07B2200/11

  Compounds covalently bound to a solid support · CPC title

• C12Q2563/107

  fluorescence · CPC title

• C12Q1/6869 Primary

  Methods for sequencing · CPC title

• C12Q1/6874

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