Enabling high-accuracy long-read amplicon sequences using unique molecular identifiers

Ryan Ziels: ‘Amplicon sequencing has emerged as an invaluable tool that allows us to study the diversity and discover novel lineages of microorganisms that have importance, all the way from biogeochemical cycling, all the way to human health’.

Ryan’s ‘Amplicon Sequencing Wishlist’: long amplicons, high accuracy, high throughput, easily accessible. Short-read seq does not give us long amplicons and is not that easily accessible.

Ryan used unique molecular identifiers (UMIs) for amplicon sequencing, which tag individual DNA molecules ‘like sample barcodes, only better’.

Ryan showed that their UMI approach improved accuracy to >Q40, or error ~0.006%. We can generate ‘almost perfect reads using this UMI tagging approach’.

Ryan pointed to B. subtilis which has 10 rRNA operons, differing by only a few nts. All 10 were successfully resolved.