Pangenome Graph Construction from Genome Alignment with Minigraph-Cactus

Reference genomes provide mapping targets and coordinate systems but introduce biases when samples under study diverge sufficiently from them. Pangenome references seek to address this by storing a representative set of diverse haplotypes and their alignment, usually as a graph. Alternate alleles determined by variant callers can be used to construct pangenome graphs, but thanks to advances in long-read sequencing, high-quality phased assemblies are becoming widely available. Constructing a pangenome graph directly from assemblies, as opposed to variant calls, leverages the graph’s ability to consistently represent variation at different scales and reduces biases introduced by reference-based variant calls. Pangenome construction in this way is equivalent to multiple genome alignment, which is extremely computationally demanding. Here we present the Minigraph-Cactus pangenome pipeline and demonstrate its ability to build a pangenome graph by aligning 90 human haplotypes. This tool was designed to build graphs with as much information as possible while still being practical for use with current mapping and genotyping tools. We show that this graph is useful both for studying variation within the input haplotypes, but also as a basis for mapping and variant calling. We also demonstrate that this method can apply to nonhuman data by showing improved mapping and variant detection sensitivity with a Drosophila melanogaster pangenome. ### Competing Interest Statement The authors have declared no competing interest.