Patent ID: 9193993
Filing Date: 2015-11-24
Classification: C12P,C12Q

Abstract:
1. A method for amplifying a target nucleic acid sequence comprising the steps of: (a) providing target probes, having a first end and a second end, that are complementary to an original target nucleic acid sequence segment wherein at least one target probe comprises at least one destabilizing moiety that is located at or near the first end that is to be ligated of the target probe; (b) hybridizing the target probes to the original target nucleic acid sequence segment and ligating the target probes together to form a first product duplex comprising the original target nucleic acid sequence segment and a newly generated destabilizing template; wherein said at least one destabilizing moiety that was located at or near the first end of the target probe in step (a) is located near a middle of the destabilizing template; (c) dissociating the first product duplex spontaneously as a result of a destabilizing effect of the destabilizing moiety to release the original target nucleic acid sequence segment and the destabilizing template; (d) providing template probes complementary to the destabilizing template that when ligated together form a product nucleic acid sequence that consists substantially of the same original target nucleic acid sequence segment; (e) hybridizing the template probes to the destabilizing template and ligating the template probes together to form a second product duplex comprising a destabilizing template and a newly generated product nucleic acid sequence that consists substantially of the same original target nucleic acid sequence segment; (f) dissociating the second product duplex spontaneously as a result of a destabilizing effect of the destabilizing moiety to produce the destabilizing template and a product nucleic acid sequence that consists substantially of the same original target nucleic acid sequence; and (g) repeating steps (a) to (f) to produce multiple copies of the destabilizing template and multiple copies of a product nucleic acid sequence that consists substantially of the same original target nucleic acid sequence segment, wherein the destabilizing moiety is selected from the group consisting of: an abasic nucleotide, a 1′,2′-dideoxyribose-5′-phosphate, butyl, cis-butenyl, or ethyl group; wherein the method is an isothermal method and the temperature of the liquid is maintained within a temperature range of about 5° C.; wherein the method for amplifying a target nucleic acid sequence is conducted in a liquid.