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natir avatar natir commented on August 24, 2024

Hi @SergeiF1987,

Thank for your issue.

First, yacrd perform two similar but different task, detection (and management of chimera) and scrubbing. If I understand you correctly you want detect chimera, but the parameter you use is designed for scrubbing not for chimera detection.

Second:
If I understand you correctly, you think the -n parameter control the length of not covered region yacrd needs to mark a read as Chimeric, it's not the case.

  • yacrd mark a read as Chimeric if a region in middle of read has coverage equal or bellow than the value in -c parameter. yacrd didn't check the length of this region if it's only one base, yacrd mark the reads as chimeric, it's not very clever. I can probably had a parameter to control the minimal length of coverage drop.
  • yacrd mark a read as NotCovered, if large part of read isn't covered, this proportion is control by parameter -n. If a read isn't covered we can't say if this read is chimeric or not and we probably can't trust him.

So if you only want detect chimera I recommend you to use this parameter:

minimap2 -x ava-ont reads.fasta reads.fasta > overlap.paf
yacrd -i overlap.paf -o report.yacrd -c 0 -n 0.8

If a read have a drop of coverage to 0 it's probably a chimera, if read isn't covered more than 80 percent of length this read can't be trust.

To improve the readme can you please indicate to me what made you think that the -n parameter controlled the size of not covered region?

Thanks in advance

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SergeiF1987 avatar SergeiF1987 commented on August 24, 2024

Hi @natir,

Thanks for such a quick reply! Actually, -n parameter looks clear, a ratio of zero-covered to a total read length. But -c... Sorry for that, but I still don't get how a region with 0 coverage could be chimeric? Does it mean that a read has no coverage at all? In this case, a sequence is presented only once and could stem from a weird source (sequencing errors, etc.) and considered as chimeric.

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natir avatar natir commented on August 24, 2024

In yacrd we assume, if part of read have no coverage this part didn't exist in other reads and probably denote this read is chimeric (or this region have very high error rate).

We use minimap2 to search overlap between reads and use this overlap to evaluate coverage.
minimap2 didn't perform exact alignment between reads, it perform overlap search, two reads share an overlap if they seem the same not exactly the same, so small sequencing errors are ignored.

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SergeiF1987 avatar SergeiF1987 commented on August 24, 2024

I get it! Thanks a lot! Perhaps you could add a more detailed explanation of this in the manual for slow minds like mine.
Thanks again and good luck!

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