QUANTITATIVE AMPLICON SEQUENCING FOR MULTIPLEXED COPY NUMBER VARIATION DETECTION AND ALLELE RATIO QUANTITATION

摘要

Provided herein are methods of quantitative amplicon sequencing, for labeling each strand of targeted genomic loci in a DNA sample with an oligonucleotide barcode sequence by polymerase chain reaction, and amplifying the genomic region(s) for high-throughput sequencing. The methods can be used for the simultaneous detection of copy number variation (CNV) in a set of genes of interest, by quantifying the frequency of extra copies of each gene. In addition, these methods provide for the quantitation of the allele ratio of different genetic identities for targeted genomic loci using multiplexed PCR. In addition, these methods provide for the detection of mutations and quantitation of the variant allele frequency.