Genetic and Protein Engineering(10 Plenary Forums - 1 Event)

Biography

Biography: Amirali Kia

Abstract

Statement of the Problem: Next generation sequencing has opened new doors into genomics. Recent advances in sequencing technology have enabled researchers to sequence genomes with unprecedented accuracy and speed. However, preparing DNA sequencing libraries still remains as a challenging step. Transposase-based library preparation off ers a simple, fast, scalable, and flexible solution. However, when compared to other (ligation-based) library preparation methods, transposases show some insertion bias that aff ects the fi nal DNA sequencing library.

Methodology: We applied various machine learning algorithms to random mutagenesis libraries of transposases to efficiently design new mutant transposases with less insertion bias.

Findings: We present the discovery of a mutant transposase (Tn5-059) with low GC insertion bias and AT dropout. Libraries prepared by this mutant have good uniformity of genome coverage as well. Tn5-059 also shows low sensitivity to the amount of input DNA. In addition, this enzyme shows effi cient performance in DNA sequencing libraries for open chromatin profiling.

Conclusion & Significance: DNA input tolerance together with superior coverage uniformity are two important factors to be considered in DNA sequencing library preparation. Tn5-059 successfully delivers on both of these aspects, which leads to less sequencing volume and lower sequencing cost. We discuss the importance of choosing a correct assay as well as the importance of filtering data based on the biology behind the assay.