US 11,713,484 B2 · Mapping the location, type and strand of damaged and/or mismatched nucleotides in double-stranded DNA

Abstract

Providing herein, among other things, is a method comprising incubating a double-stranded nucleic acid having a nick with a nick translating activity, a ligase, and a nucleotide mix comprising at least one modified nucleotide, to generate a product comprising a patch of a newly synthesized strand of a duplex nucleic acid containing a plurality of modified nucleoside monophosphates that are at or adjacent to the site of the nick. In some embodiments, the method may be used to map damaged nucleoside monophosphates in a nucleic acid. Compositions and kits for use in performing the method are also provided.

Claims34

Dependent claims

A method comprising:

nick translating a double-stranded nucleic acid comprising a nick in a reaction mix that comprises:
one or a plurality of enzymes that provide nick translating activity, a nucleotide mix comprising at least one modified nucleotide and a ligase,
wherein nick translation is initiated at the nick, a plurality of the at least one modified nucleotide is incorporated at or adjacent to the nick during nick translation and the ligase seals the nick translated nucleic acid,
wherein the at least one modified nucleotide is not fluorescent or biotinylated and/or the method involves sequencing the nick translated nucleic acid.

1 more element

Dependencies are inferred from the claim text.

Facts

Application

16/758,190Filed 21 Aug 2018 · us provisional application

Priority

16 Nov 2017

Published

US 2020/0325533 A115 Oct 2020

Granted

1 Aug 2023

Term ends

Not estimatedThe source records don’t support an estimate.

Classification8 codes

C12Q 1/6869
Involving nucleic acids: methods for sequencing
C12Q 1/6806
Involving nucleic acids: preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
C12Q 2521/101
Nucleotidyl transfering: dNA polymerase
C12Q 2521/307
Phosphoric diester hydrolysing, i.e. nuclease: single strand endonuclease

Mapping the location, type and strand of damaged and/or mismatched nucleotides in double-stranded DNA

Life of this patent

Mapping the location, type and strand of damaged and/or mismatched nucleotides in double-stranded DNA

Life of this patent

Abstract

Claims34