Latest chemistry retains the Q50 accuracy of R10 and provides increased throughput and capture, better raw read accuracy and compatibility with PromethION. Designed to improve on R10, which has recently reported at 99.995% single molecule consensus accuracy with UMI method.
Flow cells using the latest nanopore chemistry, R10.3, are now available to purchase in the nanopore store. MinION Flow Cells will ship from next week, and PromethION flow cells during February.
Constant performance iteration
Oxford Nanopore constantly upgrades the performance of its sequencing technology, by a variety of methods including introducing new types of nanopore whose different structures generate different signal properties.
The current, broadly-used nanopore, R9.4.1, has been used in multiple publications that show rapid generation of high-quality sequence data and testing, across multiple disciplines including cancer research, human genetics and microbiology. The most up to date analysis tools, such as the flip-flop basecaller and Medaka for consensus generation, support high-throughput data with raw read accuracy of 95% or consensus accuracy as high as Q40 (shown below in a Zymo mock community).
In March 2019, the next generation of nanopore – R10 – was released into early access by a small number of users. Enhanced performance was reported in 2019 by users of R10 and internal results showed consensus accuracy as high as Q50.
For example, in a preprint featuring R10 published this week by Karst et al., the team share a high-throughput, highly-accurate, long-read amplicon sequencing approach that uses unique molecular identifiers (UMIs) with R10. They obtain a mean consensus single molecule error-rate of <<0.005%. Andrew Beggs of Birmingham University reported enhanced matches with R10 on the HLA region of the genome, with a view to tissue typing. Grandomics reported improved consensus accuracy and high- accuracy variant calling in a 2Mb region associated with Duchenne’s Muscular Dystrophy using R10.
R10 has a longer barrel and dual reader head, enabling improved resolution of homopolymeric regions and improving the consensus accuracy of nanopore sequencing data.
R10.3: Higher throughput and improved accuracy
Since the introduction of R10, the Oxford Nanopore R&D teams have been working to tune the pore for even higher performance (see below), including:
- increased throughput and capture, including compatibility with PromethION flow cells
- input amounts closer to R9.4.1
- improved raw accuracy to match R9.4.1, to support enhanced variant calling
The newest R10.3 variety was introduced by Clive Brown during his technology update at NCM 2019 (31:50).
R10.3 flow cells will be available across all devices with MinION (FLO-MIN111) and PromethION (FLO-PRO111) now in store and Flongle flow cells coming later. R10.3 will be suitable for use with the LSK109 kit at launch.