Leszek
@nanopore DRS sequencing temperature was lowered to 30/31C (#P2 / #MinION) in #minknow v24.06 (and all recent versions) slowing down RNA helicase from 130 bps to 100-110 bps, lowering yield per hour and affecting signal readouts.
In effect, recent and older #RNA004 runs differ in base- and modification calling quality, possibly pA tail estimates.
This change was not widely announced to the community apart from a brief mention in software release notes!
https://community.nanoporetech.com/posts/software-release-24-06-for
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Aug 13, 2025 at 10:18amWeb
Show threadLeszekAug 13, 2025
No one in our lab was aware of this change until now. Started digging after seeing strange results when comparing runs from 2023-24 and 2025!
Show threadChristian MeestersAug 13, 2025
@lpryszcz Thank you! I will forward this information to my colleagues. Being curious whether they knew this already.
Show threadLeszekAug 13, 2025
Different helicase speed requires retraining all basecalling models. This change probably coincides with v5.0.0 models released with dorado v0.7.0
https://github.com/nanoporetech/dorado/releases/tag/v0.7.0
Because of that I suspect v5 models won't be optimal for basecalling older RNA004 runs sequenced at higher 35C / 130 bps (minknow older than v24.06, so before summer 2024).
Release v0.7.0 · nanoporetech/dorado

[0.7.0] (21 May 2024) This release of Dorado introduces new and more accurate v5 models for improved basecalling. It also adds a new subcommand, dorado correct, for single-read error correction to ...

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