Microbes play an important role in nearly every part of our world, as they affect human health, our environment, agriculture, and aid in waste management. Complete closed genome sequences, which have become the gold standard with PacBio long-read sequencing, can be key to understanding microbial functional characteristics. However, input requirements, consumables costs, and the labor required to prepare and sequence a microbial genome have in the past put PacBio sequencing out of reach for some larger projects. We have developed a multiplexed library prep approach that is simple, fast, and cost-effective, and can produce 4 to 16 closed bacterial genomes from one Sequel SMRT Cell. Additionally, we are introducing a streamlined analysis pipeline for processing multiplexed genome sequence data through de novo HGAP assembly, making the entire process easy for lab personnel to perform. Here we present the entire workflow from shearing through assembly, with times for each step. We show HGAP assembly results with single or very few contigs from bacteria from different size genomes, sequenced without or with size selection. These data illustrate the benefits and potential of the PacBio multiplexed library prep and the Sequel System for sequencing large numbers of microbial genomes.
Organization: PacBio
Year: 2018
