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Basecalling options
We offer two options for basecalling Kit 12 data with sequencing accuracies of 99% and above (Q20+):
- Simplex basecalling, where the template DNA strand passes through the nanopore and is basecalled.
- Duplex basecalling, where the complement strand is read immediately after the template strand and the consensus basecall for both strands leads to a further increase in accuracy.Each of these options is described in more detail in the "Basecalling Kit 12 data" subsections:
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We are now offering a duplex read basecaller in Guppy (duplex_tools), where the template and complement strands of a read can have their basecall data combined to provide a more accurate sequence.
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Run simplex basecalling and read splitting in MinKNOW or Guppy.
More information on this can be found in the previous section, Basecalling Kit 12 simplex data.
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Ensure you have installed Guppy version 6.0.0 or higher.
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Run duplex basecalling in Guppy.
The "duplex basecalling" section of the Guppy protocol contains instructions for how to perform duplex basecalling. To perform duplex basecalling, the template and complement read pairs must first be identified. After read pairing and filtering, the
duplex_tools
executable can then be launched.