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Require a minimum sequence length of 50bp after ITSx #718

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merged 1 commit into from
Mar 26, 2024
Merged

Require a minimum sequence length of 50bp after ITSx #718

merged 1 commit into from
Mar 26, 2024

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d4straub
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@d4straub d4straub commented Mar 25, 2024 •
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Sequences below 50bp cannot be used by DADA2 and most likely also other kmer-based classification methods fail with too short sequences (see https://github.com/benjjneb/dada2/issues/601). This can be usually easily prevented with --min_len_asv, but not when ITSx is used (because this is after --min_len_asv is appplied). Therefore, another length filtering step with 50bp min length is added here. The minimum value of 50 is not represented by a parameter but can be adjusted via a config, if required.

Closes #704.

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@d4straub d4straub requested a review from jtangrot March 25, 2024 15:26
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nf-core lint overall result: Passed ✅ ⚠️

Posted for pipeline commit f15c9dd

+| ✅ 191 tests passed       |+
#| ❔   6 tests were ignored |#
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  • nf-core/tools version 2.13.1
  • Run at 2024-03-25 15:29:11

erikrikarddaniel
erikrikarddaniel approved these changes Mar 25, 2024
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👍

@d4straub
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Thanks!

@d4straub d4straub merged commit c828ef4 into nf-core:dev Mar 26, 2024
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@d4straub d4straub deleted the fix-itsx-short-sequences branch March 26, 2024 08:01
jtangrot
jtangrot approved these changes Mar 26, 2024
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Looks good! Thanks! My only concern is that the length filtering is generally applied, and not only when running DADA2 for taxonomies. E.g. sintax by default discards sequences shorter than 32, not 50, but this can be changed with a parameter and I think there is no requirement on sequence length (not going into details on how short a sequence can be to achieve sensible results). So for sintax a sequence length of 50 is not technically needed. That being said, I guess it's easy to change the filtering cutoff in ampliseq with a custom config if one thinks 50 is too much.

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Thanks for your review! Unfortunately I seem to have merged the PR just before your comment.
I actually do not know what methods require a minimal length and which not, just DADA2 seems certain.
I considered 2 solutions: (1) adding a filter after ITSx to control its output for all downstream processes, or (2) adding a length filter before DADA2.
My impression was that short sequences are generally not advisable to classify (and didnt know that sintax has a build-in filter) and therefore I decided for option (1). Would you have preferred option (2)?

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Yes, I realized I was too slow when posting my comment :D I'm fine with this solution too.

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@jtangrot jtangrot jtangrot approved these changes

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