Comparative analysis of alignment tools for application on Nanopore sequencing data

Abstract

INTRODUCTION: Long-read sequencing techniques such as Oxford Nanopore sequencing, are representing a promising novel approach in molecular-biological methodology, enabling potential facilitation in mapping and de novo assembly. In comparison to conventional sequencing methods, novel alignment tools are mandated to compensate differing data structures (especially high error rate) to achieve acceptably accurate analysis results. METHODS: In this study, benchmarking for long read aligners BLASR, GraphMap, LAST, minimap2, NGMLR and the short-read aligner BWA MEM on three experimental datasets was conducted. Obtained alignment results were compared for various quality and performance criteria, such as match rate, mismatch rate, error rate, working memory usage and computational time. RESULTS: The comparison yielded differences in alignment quality and performance of tools under test. Tool LAST showed the largest differences among all tools. Minimap2 achieved constant quality with good performance. BLASR, GraphMap, BWA MEM and NGMLR showed slight differences only. CONCLUSION: Differences among the tools could be reasoned with dataset characteristics and algorithm approaches of individual tools. All tools except BLASR seem applicable for Nanopore sequencing data. Therefore, selection of the tool should be done under consideration of the experimental design and the further downstream analysis

Keywords

• alignment tools
• nanopore sequencing
• benchmarking
• comparative analysis