Western Sydney University, Australia
Title: Enhancing NGS performance through improvements in template preparation procedure
Biography: Caroline Janitz
Statement of the Problem: Over the past few years next-generation sequencing (NGS) technology has been broadly implemented across diverse research areas. Despite increasing sequencing throughput, sample preparation still remains a technical challenge. Slight modifications to the template preparation protocol may improve the quality of sequencing data and lead to a reduction in sequencing costs. We will present a number of improvements in template preparation protocol with examples from a variety of NGS applications.In metagenomics projects, un-normalised input DNA from different treatments can significantly affect sequencing outcomes at species level leading to a biased diversity estimate.
The main challenge for epigenetic NGS projects is a relatively high duplicate rate which results from the necessity for an increased number of PCR cycles frequently required to obtain enough material for sequencing. A simple reduction in the number of unnecessary PCR cycles can significantly diminish the duplicate rate resulting in enhanced ChIP-Seq data quality. Strand-specific RNA-Seq has been widely implemented in the field of transcriptomics, although template preparation still remains challenging, particularly for clinical samples. We will demonstrate that the implementation of minor improvements to the template preparation protocol results in dramatic amelioration in the quality of sequencing output