A fundamental requirement for genetic studies is an accurate determination of sequence variation. While human genome sequence diversity is increasingly well characterized, there is a need for efficient ways to use this knowledge in sequence analysis. Here we present Graphtyper, a publicly available novel algorithm and software for discovering and genotyping sequence variants. Graphtyper realigns short-read sequence data to a pangenome, a variation-aware graph structure that encodes sequence variation within a population by representing possible haplotypes as graph paths. Our results show that Graphtyper is fast, highly scalable, and provides sensitive and accurate genotype calls. Graphtyper genotyped 89.4 million sequence variants in the whole genomes of 28,075 Icelanders using less than 100,000 CPU days, including detailed genotyping of six human leukocyte antigen (HLA) genes. We show that Graphtyper is a valuable tool in characterizing sequence variation in both small and population-scale sequencing studies.
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We are grateful to our colleagues from deCODE Genetics/Amgen for their contributions. We also wish to thank all research participants who provided biological samples to deCODE Genetics.
All authors are employees of deCODE Genetics/Amgen, Inc.
Integrated supplementary information
Supplementary Figure 1 IGV visualization of mapped sequence reads of two Icelanders carrying a 40-bp deletion.
The genomic region shown is chr. 21: 21,559,430–21,559,518 (GRCh38), and the deleted sequence is between the two vertical green lines. (a) A heterozygous carrier of the deletion. Graphtyper was the only genotyping pipeline that correctly recognized the individual as a carrier. The other pipelines called the false sequence variants due to misalignments around the indel (red boxes). (b) A homozygous carrier of the deletion. In this case, most of the reads are correctly mapped as carrying the deletion, but again some misalignment artifacts are observed (red box).
Supplementary Figure 2 Alternative allele transmission rate in 230 Icelandic parent–offspring trios.
All genotyping pipelines have an excess of sequence variants that are never transmitted from parent to offspring, which may be calls due to sequencing error or non-germline variation. Bin width is 0.1.
Supplementary Figure 3 Alternative allele transmission rate in 230 Icelandic parent–offspring trios by SNP mutation type.
The mutation ratio of transitions and transversions is estimated to be around two in the human autosomal genome. We observed that the transition/transversion ratio improved at higher transmission rates, indicating that transmission rate is measuring quality. There was also a large excess of transversions that are not transmitted.
(a) Semi-global banded alignment of a sequenced read to an extracted reference sequence. (b) Observed variation with respect to the reference sequence.
(a) An example of a graph with two sequence variants. The graph has a total of six haplotypes. (b) Two sequence variants are closer than 5 bp to each other and are grouped together. (c) If we only observed four of six haplotypes in a population, we can reduce the number of haplotypes in the graph to four as shown here.
(a) Both parents are homozygous and the offspring’s genotype can be inferred. (b) At least one parent is heterozygous and we cannot uniquely infer the offspring’s genotype. We can measure the transmission rate of an allele in these types of trios.
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Eggertsson, H., Jonsson, H., Kristmundsdottir, S. et al. Graphtyper enables population-scale genotyping using pangenome graphs. Nat Genet 49, 1654–1660 (2017). https://doi.org/10.1038/ng.3964
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