Patent ID: 8741564
Filing Date: 2014-06-03
Classification: C12Q

Abstract:
1. A method of determining a sequence of at least one target nucleic acid molecule in a sample, comprising: contacting the sample with at least one nuclease protection probe comprising a flanking sequence (NPPF) under conditions sufficient for the NPPF to specifically bind to the target nucleic acid molecule, wherein the NPPF comprises: contacting the sample with a nucleic acid molecule comprising a sequence complementary to the flanking sequence (CFS) under conditions sufficient for the flanking sequence to specifically bind to the CFS; contacting the sample with a nuclease specific for single-stranded nucleic acid molecules under conditions sufficient to remove unbound nucleic acid molecules, thereby generating a digested sample comprising NPPFs hybridized to the target nucleic acid molecule and to the CFS(s); amplifying NPPFs in the digested sample with the amplification primer, thereby generating NPPF amplicons; and sequencing at least a portion of the NPPF amplicons, thereby determining the sequence of the at least one target nucleic acid molecule in the sample.