US 9,005,933 B2 · Helicase dependent isothermal amplification using nicking enzymes

Abstract

The present invention relates to a method for amplifying a template nucleic acid, wherein the method comprises amplifying said template nucleic acid using the helicase dependent amplification (HDA) reaction in the presence of a nicking endonuclease, and wherein said template nucleic acid comprises a sequence recognized by said nicking endonuclease or a sequence recognized by said nicking endonuclease is introduced into the template nucleic acid during the HDA reaction. The invention further pertains to a kit for amplifying a nucleic acid, comprising a nicking endonuclease, a helicase and a DNA polymerase.

Claims19

Dependent claims

A method for amplifying a template nucleic acid, comprising:

(a) providing a template nucleic acid,
(b) contacting the template nucleic acid with a helicase,
(c) hybridizing an oligonucleotide primer to the template nucleic acid, and
(d) amplifying said template nucleic acid using the helicase dependent amplification (HDA) reaction, said HDA reaction introducing a recognition sequence into the template nucleic acid,
said HDA reaction occurring in the presence of a nicking endonuclease that recognizes a nucleotide sequence of a completely or partially double-stranded nucleic acid molecule and cleaves only one strand of the template nucleic acid at a specific location relative to the recognition sequence,
wherein the HDA reaction is a thermophilic HDA (tHDA),
and wherein the template nucleic acid is shorter than 400 bp.

4 more elements

Dependencies are inferred from the claim text.

Helicase dependent isothermal amplification using nicking enzymes

Life of this patent

Helicase dependent isothermal amplification using nicking enzymes

Life of this patent

Abstract

Claims19