Fully phased human genome assembly without parental data using single-cell strand sequencing and long reads.

Porubsky, David; Ebert, Peter; Audano, Peter A; Vollger, Mitchell R; Harvey, William T; Marijon, Pierre; Ebler, Jana; Munson, Katherine M et al. · Nat Biotechnol · 2021

basic_science · Level V

Where this comes from

Abstract

Human genomes are typically assembled as consensus sequences that lack information on parental haplotypes. Here we describe a reference-free workflow for diploid de novo genome assembly that combines the chromosome-wide phasing and scaffolding capabilities of single-cell strand sequencing<sup>1,2</sup> with continuous long-read or high-fidelity<sup>3</sup> sequencing data. Employing this strategy, we produced a completely phased de novo genome assembly for each haplotype of an individual of Puerto Rican descent (HG00733) in the absence of parental data. The assemblies are accurate (quality value > 40) and highly contiguous (contig N50 > 23 Mbp) with low switch error rates (0.17%), providing fully phased single-nucleotide variants, indels and structural variants. A comparison of Oxford Nanopore Technologies and Pacific Biosciences phased assemblies identified 154 regions that are preferential sites of contig breaks, irrespective of sequencing technology or phasing algorithms.

Medical subject headings