COMPOSITIONS AND METHODS FOR DETECTING RARE NUCLEIC ACID MOLECULE MUTATIONS

摘要

The present disclosure relates to compositions and methods for detecting rare nucleic acid molecule mutations in a plurality of nucleic acid molecules. Also disclosed are methods for determining the size of a nucleic acid molecule using droplet digital PCR.

主权项

1. A method for detecting rare nucleic acid molecule mutations, comprising:
a. contacting a plurality of nucleic acid molecules with a first restriction endonuclease, wherein the first restriction endonuclease is capable of cleaving a nucleic acid molecule comprising a first target region having a site specific for the first restriction endonuclease, and wherein the nucleic acid molecule is not cleaved by the first restriction endonuclease when the first target region comprises a mutation that alters the site specific for the first restriction endonuclease; b. amplifying the mutated first target region from the plurality of nucleic acid molecules of step (a) with a first 5′ primer and a first 3′ primer, wherein the primers are complementary to nucleic acid sequences flanking the first target region on a nucleic acid molecule, and wherein only the mutated first target region is substantially amplified; and c. quantifying the amplified amount of mutated first target region, thereby detecting a rare mutation within a plurality of nucleic acid molecules.