Patent ID: 9689031
Date: 2017-06-27
CPC Classifications: C07H,C12Q,G01N

Claim:
1. A method for amplifying a double-stranded nucleic acid target sequence, comprising: a) contacting a target DNA molecule comprising a double-stranded target sequence having a sense strand and an antisense strand, without, prior to the contacting, subjecting the target DNA molecule to a thermal denaturation step associated with amplification of the target sequence, with: (1) a forward template and a reverse template, wherein (2) a first nicking enzyme that is capable of nicking at the nicking site of said forward template, and does not nick within said target sequence; (3) a second nicking enzyme that is capable of nicking at the nicking site of said reverse template and does not nick within said target sequence; (4) a DNA polymerase; and b) amplifying the double-stranded nucleic acid target sequence under essentially isothermal conditions; wherein amplification is performed by multiple cycles of said polymerase extending said forward and reverse templates along said target sequence producing a double-stranded nicking site, and said nicking enzymes nicking at said nicking sites, and, following a first round of amplification, at amplified copies of said sites, producing an amplification product.