Oxford Nanopore releases ‘Rev D’ flow cells, enabling increase in data yields

Oxford Nanopore has released a new version of MinION and GridION flow cells that include the new ‘Rev D’ ASIC. The new release extends the amount of time that flow cells can be used for DNA sequencing or RNA sequencing, increasing the overall yields of DNA sequence data to as much as 30 Gb per flow cell (at this performance, the equivalent of ~10X human genome for $500*).

The amount of sequence data that can be generated with a single flow cell has increased dramatically since the launch of nanopore sequencing technology. Previous increases in yield have been driven by various elements of the technology including:

  • increasing the rate at which DNA fragments pass through a nanopore from 35 bases per second at launch to 450 bases per second now
  • the release of updated versions of MinKNOW.  These software releases deployed features including ‘progressive unblock’ and ‘active voltage control’
  • the release of ‘Kit 9’, which maximises both read length and yields in a single kit.

With no capital investment required for devices and as consumable prices have remained consistent since launch, this translates into substantial cost-per-base decrease for users of nanopore sequencing (for more details, review our spec table).

PromethION for ultra-high throughput, long-read sequencing

PromethION is based on a different ASIC; each flow cell contains 3,000 channels and can now yield beyond 130 Gb in the field and as much as 200 Gb in internal use. Each PromethION is designed to run up to 48 flow cells at any time.  At $625 per flow cell for the largest consumables package and with no capital investment required for the device, this enables a low-cost, long-read human genome.

Read length = input fragment length

Nanopores process the entire DNA or RNA fragments in the sample; read length is dependent on the method of sample preparation chosen by the researcher. In comparison to circular sequencing methods, nanopore read lengths are the full fragment length seen in the sample, rather than the sum of multiple repeat passes of a smaller fragment. Nanopore users commonly report a ‘read N50’ of over 30kb.  Read N50 refers to a value where half of the data is contained within reads with alignable lengths greater than this.  With typical (non targeted) sample preparation methods, nanopore reads will be dispersed evenly across the genome for uniform coverage.

The portable MinION runs one flow cell and desktop GridION can run up to five flow cells at any time. As nanopore sequencing streams data in real time, flow cells can be run until sufficient data has been collected and then stopped; they can then be stored, washed and used again later.