Albert Vilella Profile picture
May 19, 2022 57 tweets 16 min read Read on X
The London stock market is now closed and $ONT.L Oxford @nanopore is presenting technology updates. I'll live tweet my highlights, but won't be thoroughly complete with screenshots (thread)
My COIs bit.ly/avilcoi
First talk is James Clark for Platform updates.
But Clive is already saying there will be a "thing at the end" that you want to stick around for. Reminiscent to the "one more thing" that certain Apple presentations had for a while.
Current platform with the P2 released in 2022
Flowcell pore counts are 1x to 4x to 24x from Flongle to MinION to PromethION flowcells. So 3 main "platform sizes", similar to the 3 main platform sizes (instruments) that $ILMN Illumina has.
PromethION best of all flowcell now, latest iteration of ASIC and the rest. P2 instrument makes it more accessible for all budgets.
Chemical and sample loading changes to optimise the process
The triangle of better, cheaper and faster, here Quality, Volume and Features. Biosensing continuous iteration process.
New products coming soon, mainly new ASIC-derived in the near future.
New ASIC being road tested on Flongle flowcells
New ASIC has a simpler printing, small path to the electrodes, which means high accuracy.
My take from here: the ASIC part of the technology is akin to the physics/electronics part of other sequencing technologies. The rest is molecular biology, similar to the polymerases and reagents in other technologies. Both bound by our still incomplete understanding of both.
Nanopore Chemistry. Where are we and where are we going?
The short blanket problem of optimising both throughput and accuracy, here displayed in an x-y plot over time
Accuracy plots and pore speed plots of current iteration
This is tunable: faster or more accurate in different modes.
It's already out and people are testing it in the field. Smoothest Early Access ever.
Duplex reads, let's see what is the rate of efficiency of the process, going up to 40%, with a target of 60%. If we compare to Illumina's Paired End, that would seem to be 100% but...
... remember Illumina only shows the successfully paired-end reads, so it has to be 100% by definition.
The new pore is a nonamer (reads 9-nts) and is what has taken simplex accuracy to 98%.
Better Simplex, better Duplex, overall better accuracy. What else can be improved? Homopolymer chemistry improvements.
How do we define accuracy? Raw read, variant calling, consensus, testing accuracy, etc.
On test accuracy, nothing like COVID19 variant surveillance to describe what the technology can do, both in terms of accuracy but in Oxford @nanopore's case also in ease of deployment.
Duplex accuracy very independent of read length. $PACB PacBio engineers may look at this and be a bit worried that Oxford @nanopore has something better than HiFi.
Nothing like an IGV @igvteam plot to show how things have progressed
Consensus/assembly numbers: maybe everything will be assembly-based in human genomics in the future (as people like @lh3lh3 keep saying), so these numbers matter.
Variants, not showing DeepVariant here (not invented here bias?), but I would say in many ways on par with $PACB and $ILMN on this front.
Bsae modifications, I am particularly proud to see that 5hmC is now a first-class mod. Very important for #LiquidBiopsy early cancer screening -- people will find out soon.
Headline slide
Built-in short fragment mode, meaning getting 250M reads from a single PromethION flowcell, regardless of length.
Adaptive sampling: nobody else can do this natively. Average 3x fold enrichment, but improvements coming. Still, 3x cheaper/better than not doing it.
More formats! Pod5 format as a replacement to Fast5. Many workaround coming from feedback from the Slow5 academic project. Between 4-10x improvement in performance.
New version of the basecaller, Dorado with ground-up rewrite. I am sure Chris @iiSeymour and the team are very proud of it.
Products section by @RosemaryDokos with highlights including starter packs that make this accessible to people...
and key openness to key features to enhance the inventiveness of the users. Open and competitive pricing.
Most platform updates are *not* instrument updates, which means the user can amortize the CAPEX easily. OPEX options as well.
Release phases, ending in Q-Line and now working on CE/IVD products
Sample prep: more for every task, but also "Extraction central" to share/discuss. Including fragment length tuning sample prep methods.
Single cell methods and also software to analyse the results. Key area in my opinion. Direct RNA also improved (input amount, cleanups).
Key example is COVID19, deployed in more than 100 countries. Best example one can think of the ease of deployment feature of Oxford @nanopore "sequence everywhere" mantra.
Kit 14 ligation native barcoding at 24x and 96x, now working on 384-plex. Cas9 following shortly. Rapid as well.
RAD chemistry will go into lower inputs and Q20+ in Q3.
Slide showing the wave of Kit14 over the whole prep portfolio. Formal release aiming at Q4 2022.
The Kit14 is improving the numbers all over.
PromethION now starts at $10-11K with the P2. Potential customers running out of excuses here.
Yes but what about sequencing factories? Also happening. This is the cash cow of $ILMN Illumina currently, being threatened from multiple angles.
Biggest EPI2ME annoucement, for me, is the Nextflow integration. Great news in my opinion.
Clive is back for "one more thing"...
Actually a litany of R&D projects, all very cool, I would highlight the toothbrush continuous epigenomic profiling idea. I called this the expresso machine equivalent in the past.
Protein sequencing: this goes to the heart of offerings by companies like $QSI Quantum-SI and Erysion. Taking a go at it here: 20 aminoacid alphabet but also post-translational modifications (PTMs).
Amino calling equivalent to the basecalling in nucleobases. Combinatorics in proteins are enormous. In some ways the bar is lower than DNA sequencing, in some ways is higher.
Very reproducible profiles: looking at slight shifts in the squiggle, but the information is there.
This is far from a final product, but this approach aims at shotgun #proteomics
Democratizing proteomics for any biologist out there. Complex proteomics for all. That's the target, significant proof of concept done, still a way to go right now.
Re-reading capabilities, which are very important in proteomics where, same as RNA-seq, the distribution can be very loaded: lots of copies of boring proteins, and needle in a haystack problem for interesting ones.
There you are! Q&A now. From my part, I'll ask you to please individually 'like' the tweets in this thread that you want to highlight. What's your best bits from this update?

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More from @AlbertVilella

May 18, 2023
The tech update from Oxford @nanopore #NanoporeConf now ongoing with @The__Taybor now presenting:
Duplex: 1.5% of the time, the complement strand follows the first strand naturally.
Initially, modified the adapters and reached 30% duplex rate. Image
Stereo base caller uses similar ML approaches as ChatGPT, Image
Read 9 tweets
May 17, 2023
On library prep at #Nanoporeconf, a description for PCR-free methods showing the difference between ligation (max output) and rapid mode (10minutes, minimal lab equipment needed). Ultralong reads (ULR) also enabled, all Kit14. Image
Rapid ULR. Current record is about 4 megabases. Image
PCR expansion kits enable the use of samples with low input amount. Image
Read 10 tweets
Apr 21, 2023
I did a deep dive on the different workflow management (WFM) tools for #Bioinformatics Data Analysis a few years ago, and since then there have been a few extra entrants in this segment, still mostly concentrated in serving the Next-Generation Sequencing field.
A few years ago, there were two communities dominating the open-source WFM ecosystem in NextFlow and SnakeMake, and two platforms dominating the the commercial offerings in DNAnexus and Illumina BaseSpace.
Since then, a company out of the founders of Nextflow has started offering enterprise support for Nextflow workflows in the cloud: Seqera Labs. They offer the extra level of support that some organizations require to run Nextflow on their Data Analysis setups.
Read 7 tweets
Apr 21, 2023
More interesting Next-Generation Sequencing knowledge in the ASeq Discord channel (by @new299). Illumina patterned flowcells and the etching process to "print" the wells into the flowcell. Could be down to 350nm diameter for some flowcell configurations now. Image
If I remember correctly, Illumina started with a 600nm diameter for the patterned flowcell, in the HiSeq X and then later on in the evolution of the platform that used these patterned flowcells.
They then said to have gone down to 500nm, and what you are showing seems to indicate that it's at 350nm now, at least for the NextSeq 2000? I am not sure if they claimed that for NovaSeq X?
Read 8 tweets
Apr 20, 2023
There have been some acquisitions in #CancerDiagnostics and #CancerScreening recently, some of which signify a trend towards consolidation that is worth describing:
$A Agilent is moving towards some more vertical integration in Cancer Dx and Cancer screening
by recently acquiring both announcing a partnership with Akoya Bio and announcing the acquisition of Avida Biomed. Image
Some may ask: isn’t $A Agilent too small to go into this field? Would they be able to compete against $ILMN Illumina/GrailBio or $GH Guardant Health or $EXAS Exact Sciences?
Read 4 tweets
Apr 20, 2023
It is likely that as Spatial Biology tools become more robust and user-friendly, they will become increasingly popular and widely adopted in the scientific community.
This may lead to a shift in the balance between single-cell and Spatial Biology approaches, with the latter eventually becoming more prevalent.
Additionally, as more and more datasets are generated using Spatial Biology techniques, the field of Machine Learning and Artificial Intelligence will likely play an increasingly important role in analysing and interpreting this data.
Read 6 tweets

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