Applying Sequel to Genomic Datasets
De novo assembly is a large part of JGI’s analysis portfolio. Repetitive DNA sequences are abundant in a wide range of organisms we sequence and pose a significant technical challenge for assembly. We are interested in long read technologies capable of spanning genomic repeats to produce better assemblies. We currently have three RS II and two Sequel PacBio machines. RS II machines are primarily used for fungal and microbial genome assembly as well as synthetic biology validation. Between microbes and fungi we produce hundreds of PacBio libraries a year and for throughput reasons the vast majority of these are >10 kb AMPure libraries. Throughput for RS II is about 1 Gb per SMRT Cell. This is ideal for microbial sized genomes but can be costly and labor intensive for larger projects which require multiple cells. JGI was an early access site for Sequel and began testing with real samples in January 2016. During that time we’ve had the opportunity to sequence microbes, fungi, metagenomes, and plants. Here we present our experience over the last 18 months using the Sequel platform and provide comparisons with RS II results.