Patent ID: 8828663
Filing Date: 2014-09-09
Classification: B01L,C12Q,G01N

Abstract:
1. A method for highly sensitive and specific detection of mutant alleles in a high background of wild-type alleles, comprising a) obtaining an assay composition comprising first polynucleotides encoding a mutant allele of a gene and second polynucleotides encoding a wild-type allele of the gene, wherein the ratio of first polynucleotides to second polynucleotides is 1:1×10 b) distributing the assay composition, or a portion thereof, into at least 10,000 aliquots, thereby producing a subset of aliquots in which the ratio of first polynucleotides to the second polynucleotides is greater than the ratio of first polynucleotides to second polynucleotides in said assay composition, wherein some aliquots comprise a first polynucleotide and essentially all aliquots comprise at least one second polynucleotide; c) combining the assay composition with amplification reagents prior to step (b) or combining the at least 10,000 aliquots with amplification reagents after step (b), wherein said amplification reagents: d) amplifying the at least a portion of the mutant allele encoded in the first polynucleotides; and e) detecting the presence or absence of amplification in each aliquot.