Patent ID: 8980563
Filing Date: 2015-03-17
Classification: C12Q

Abstract:
1. A method for amplifying target sequences within a nucleic acid sample, the method comprising: fragmenting the nucleic acid sample with a restriction enzyme, thereby producing a fragmented nucleic acid sample comprising one or more target sequences; adding circularization probes to the fragmented nucleic acid sample, wherein each circularization probe comprises a first probe region and a second probe region, wherein the first and second probe regions are respectively complementary to a first target region and a second target region of one of the target sequences, and wherein the circularization probes hybridize with target sequences to form probe-target structures in which each target sequence includes a flap of two or more bases at the 5′ end and a flap of one base at the 3′ end; removing the 5′ flaps with a 5′ flap nuclease, thereby generating probe-target structures in which each target sequence includes a 5′ end juxtaposed with the 3′ end, with the 5′ end and the 3′ end separated by a nick; ligating the 5′ ends to the juxtaposed 3′ ends of the target sequences to circularize the target sequences; and amplifying the circularized target sequences.