Investigating the effects of different library preparation protocols on STR sequencing

Abstract

Next Generation Sequencing (NGS) is transforming the landscape of the Short Tandem Repeat (STR) genotyping. NGS determines the length as well as the sequence of each allele, identifies polymorphisms in the repeat or adjacent DNA regions, allows for a greater degree of multiplexing, and generates Gigabases of reads in a single run. An additional step introduced into the DNA typing process with NGS methods is library preparation. The PCR amplicons of the targeted loci are further purified and modified with adapters and sample specific indices prior to sequencing. In this study, two PCR purification methods were used to cleanup amplification reactions prior to library construction: column-based and bead-based technologies. The influence of different PCR purification protocols on the dropout events was examined.