Reducing reference bias using multiple population reference genomes

Most sequencing data analyses start by aligning sequencing reads to a linear reference genome. But failure to account for genetic variation causes reference bias and confounding of results downstream. Other approaches replace the linear reference with structures like graphs that can include genetic variation, incurring major computational overhead. We propose the “reference flow” alignment method that uses information from multiple population reference genomes to improve alignment accuracy and reduce reference bias. Compared to the graph aligner vg, reference flow exhibits a similar level of accuracy and bias avoidance, but with 13% of the memory footprint and 6 times the speed.