NCM 2019 technology update from Clive Brown
Fri 6th December 2019
Introduction - Clive recaps on how nanopore sequencing works.
Sequencing at scale (6:44) - PromethION is a new model for high throughput sequencing. Each of its 48 flow cells can be used on-demand, and if in a core lab then different users can share the device. "It's a genome centre in a box."
We’re now increasingly focusing on end-to-end high throughput workflows. Introducing XL kits for PromethION, which can prep 48 libraries, compatible with automation. Next: XL wash kit, integrating nuclease wash.
Sequencing for everyone (10:00) - Updates on Flongle, Mk1C, VolTRAX.
Not just long read (14:44) - A nanopore will process DNA or RNA, whatever the length. While nanopore is best known for long reads, the tech can sequence any fragment length.
Multiple applications that use short fragments can benefit from nanopore features: fast, cheap, modified bases, portable, real-time. Clive discusses the nanopore approach to shorter fragments.
Pushing the accuracy (19:52) - Raw read accuracy: 1D DNA basecalling is now always performed using flip-flop, which is integrated in MinKNOW. High accuracy 1D basecaller provides ~95% modal raw accuracy. Direct RNA is now comparable in performance.
Clive introduces a new basecaller, Bonito. This is elegant, using a relatively small amount of code, making the whole package much more accessible to people who want to train their own basecallers.
Chemistries for improved accuracy (31:50) - Clive introduces new variants of the R10 pore and describes methods in development to enhance single molecule accuracy method.
Skunkworkz (41:44) - Clive reviews droplet based sequencing on VolTRAX – to enable prep and sequencing of DNA, RNA or proteins on one device. And some other new ideas...