AVITI library prep miniaturization and combining with Illumina data for phylogenomic and population genomic analyses

Abstract Recent advancements in next generation sequencing approaches allow for expansion of evolutionary research into the discovery of genetic patterns and processes underlying diversification across scales. The increased popularity of the Element Bioscience AVITI platform, partially due to the high sequencing accuracy and low cost of reagents, is becoming a viable alternative approach for generating massive amounts of comparative sequencing data across diverse organismal lineages. Using a data set of five accessions from the monocot genus Costus , we tested miniaturization conditions for generating robust, cost-effective libraries and made comparisons of data generated by AVITI and Illumina sequencing platforms to investigate the potential for combining data for population genomic and phylogenomic analyses. Our results show that the AVITI and Illumina data sets are highly congruent in terms of inferring overlapping SNPs, with only a small fraction picked up by only one of the two platforms. The rates of duplication in miniaturized libraries were much higher than in full volume libraries and in the Illumina libraries, resulting in missing SNPs and less sequence coverage when volumes are reduced. For all generated libraries, most downstream evolutionary analyses, including clustering algorithms (such as PCA) and phylogenetic inference, yielded similar results. However, Structure analyses were less consistent across datasets, with data from the most miniaturized libraries being assigned to the wrong clusters. The AVITI platform should be seen as a cost-effective approach for generating genomic data for comparison across taxonomic lineages, even for ongoing projects where Illumina data already exists.