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Scalable and rare-variant aware genome inference across the 1kGP cohort

Ebler, J.; Prodanov, T.; Blair, A.; Lee, S. K.; Ebert, P.; Human Pangenome Reference Consortium, ; Paten, B.; Marschall, T.

2026-07-03 bioinformatics
10.64898/2026.06.29.735275 bioRxiv
Show abstract

Pangenome graphs built from haplotype-resolved de novo assemblies enable accurate analysis of genetic variation. The short-read-based tool PanGenie efficiently genotypes variants discovered in a pangenome across large cohorts and outperforms linear reference-based methods for structural variants (SVs). However, it cannot detect novel variants absent from the graph, missing many rare SVs (allele frequency <1%) and was limited to graphs with 254 haplotypes. First, we introduce a haplotype sampling step that reduces the number of haplotypes using sample-specific k-mers before genotyping, decreasing runtime twelvefold and memory usage 1.4-fold at 30x coverage. Second, we present a polishing workflow that corrects residual errors in haplotypes inferred from PanGenie genotypes and incorporates rare and private mutations. We genotype 3,202 samples from the 1000 Genomes Project and use low-coverage ONT data (967 samples) for polishing. We achieve a median QV of 46 and provide the 1,934 polished haplotype sequences as a community resource.

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