Tonight I continued watching the PAG Industry Workshop offered by Oxford Nanopore Technologies. Sean Prosser from the University of Guelph presented “From pandemic preparedness to planetary biodiversity: ONT enables targeted amplicon sequencing at massive scale.” Prosser noted that there are almost 35,000 amplicon-based genetic tests available in the US per year. Prosser also noted that there are 18.1 million “dark taxa” and DNA barcoding can be used. A 658 bp CO1 gene can be amplified to learn about biodiversity. Prosser and team are surveying globally through partners, including school children. As sequencing platforms have evolved, the team has used Sanger sequencing and later PacBio. With Oxford Nanopore Technologies (ONT), samples can be pooled and sequenced. Samples can be tagged using barcodes with PCR. A dual tag system can be used. For example, 96 samples can be pooled and then native barcodes are added. These samples had already been sequenced with the PacBio Sequel. Samples sequenced with ONT were comparable to those previously sequenced with the PacBio. The team then tried to sequence a million samples using dual-ligation of native barcodes with the pooled libraries. Using a PromethION flow cell, the final library was sequenced. The data has been challenging to work with with 2 Tb per run. Prosser is excited about the possibilities of using this approach to sequence many more samples!
