METHOD OF CORRECTING AMPLIFICATION BIAS IN AMPLICON SEQUENCING

摘要

A method to correct amplification bias in amplicon sequencing is disclosed. Amplification efficiency is not constant among different loci in a sample, nor for the same locus in different samples. Differences in 3'-end stability, primer Tm, amplicon length, amplicon GC content, and GC content of amplicon flanking regions all may contribute to amplification bias. Such bias interferes with accurate calculation of copy number for a genomic region of interest and hinders the application of amplicon sequencing for detection of minor copy number variation. The methods allow correction of amplification bias and enable detection of minor copy number variation using amplicon sequence data.