Benchmarking Low-Frequency Variant Calling With Long-Read Data on Mitochondrial DNA.

ABSTRACT: Background: Sequencing quality has improved over the last decade for long-reads, allowing for more accurate detection of somatic low-frequency variants. In this study, we used mixtures of mitochondrial samples with different haplogroups (i.e., a specific set of mitochondrial variants) to investigate the applicability of nanopore sequencing for low-frequency single nucleotide variant detection. Methods: We investigated the impact of base-calling, alignment/mapping, quality control steps, and variant calling by comparing the results to a previously derived short-read gold standard generated on the Illumina NextSeq. For nanopore sequencing, six mixtures of four different haplotypes were prepared, allowing us to reliably check for expected variants at the predefined 5%, 2%, and 1

SUBMITTER: Luth T 

PROVIDER: S-EPMC9161029 | biostudies-literature | 2022

REPOSITORIES: biostudies-literature