We've put out a short preprint today showing some updates in Readfish for @nanopore adaptive sampling. biorxiv.org/content/10.110… @alexomics is giving a talk shortly at the #nanoporeconf about this - but a short thread here.
Readfish now exploits the barcoding features ready baked in to @nanopore guppy to allow a few new features. You can switch off barcodes as you wish but more interestingly, you can assign different targets to different barcoded samples.
This is interesting in lots of places, but we show the use of three different human gene panels on three different samples on a single GridION flow cell.
This allows the identification of structural variants that occur between genes within the panels of interest - but it can also allow you to look at copy number variation which matches other approaches such as Bionano.
Of course - you can watch all this in real-time using our minoTour tool developed by @Rorymatics - though this is a branch currently under active development. academic.oup.com/bioinformatics…
For the real old hands who remember read until 2016 style... this allowed us to revisit an old challenge about balancing coverage over viral genomes and target individual amplicons (not in this preprint).
Can you track coverage over dozens of barcodes in real time? Well thanks to @DrT1973 and a new tool coming soon called SwordFish - we know we can! Now this does act more as a filter than enrichment but...
we do find that we can recover samples faster and recover more complete genomes by using adaptive sampling tracking 96 barcodes simultaneously. These data will be added to biorxiv.org/content/10.110… fairly soon.
As ever a lot of work by @alexomics and @Rorymatics who have both been through the @nottm_bbsrc_dtp program as iCASE students - and we happen to have an opening for another if you are interested! nottingham.ac.uk/bbdtp/case-202…

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More from @mattloose

17 Sep
Earlier this week we put out a couple of preprints (doi.org/10.1101/2021.0… doi.org/10.1101/2021.0…) describing minoTour github.com/LooseLab/minot…, an @nanopore tool we have been developing for several years. A bit of a 🧵
The minoTour design philosophy is all around real time - it's about working out what is happening at that moment - with the exception of ARTIC (see below) we anticipate users running further analysis after minoTour!
What is minoTour? It's a real time platform for monitoring and analysing nanopore data. We use it routinely in the lab to track everything we sequence (and flow cell QC too!). It's main day-to-day use is just keeping track of whats happening.
Read 15 tweets
8 Sep
An ultra-long read update. Since @circulomics is "no longer available" for @nanopore people might be interested in these protocols from @longreadclub @NininUon and @DrT1973 - a toolkit of options here dx.doi.org/10.17504/proto… including monarch from @NEBiolabs - 🧵below with more.
Ultra-long sequencing (N50s >100kb) jumped in performance in 2020 with new kits from @nanopore and @circulomics giving yields in excess of 20 Gb. In particular massively improved occupancy seemed to be a major benefit.
However, it's not always possible to start with a specific extraction method and we wanted to explore alternative methods that would give the same or better performance, using as many different approaches as possible. Little did we know that this might become more useful now!
Read 12 tweets

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