Method of amplifying nucleic acid sequences

摘要

The invention is directed to methods of removing amplicons of non target and/or target nucleic acid sequences having one or more modified (e.g., methylated) nucleotides from a sample wherein the sample comprises the non target nucleic acid and a target nucleic acid sequence to be amplified.

主权项

1. A method of removing amplicons of a non target nucleic acid having one or more methylated cytosines from a sample wherein the sample comprises amplicons of the non target nucleic acid and genomic DNA that includes a target nucleic acid to be amplified, the target nucleic acid having one or more methylated cytosines, comprising:
a) contacting the sample with a composition comprising a methyl specific restriction enzyme that specifically cleaves a nucleic acid having one or more methylated cytosines, wherein the enzyme does not cleave an unmethylated cytosine and is capable of being deactivated, thereby producing a combination; b) maintaining the combination under conditions in which the amplicons of the non target nucleic acid are digested by the methyl specific restriction enzyme prior to amplification of the genomic DNA that includes the target nucleic acid; and c) amplifying the genomic DNA that includes the target nucleic acid thereby producing amplicons of the target nucleic acid,wherein the amplicons of the non target nucleic acid are removed from the sample comprising the genomic DNA that includes the target nucleic acid to be amplified.