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ERROR: max count before zero is less than min required count (4) duplicates removed #59

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didillysquat opened this issue Feb 15, 2022 · 2 comments

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@didillysquat
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Running preseq from the latest release preseq-3.1.2.tar.gz

lc_extrap give the following error:

Singularity> preseq lc_extrap -P scaf1.1m.bam
ERROR:	max count before zero is less than min required count (4) duplicates removed

However, converting the .bam to .bed first and then running lc_extrap on the .bed file completes as expected:

Singularity> bedtools bamtobed -i scaf1.1m.bam > scaf1.1m.bed
Singularity> preseq lc_extrap -P scaf1.1m.bed > lc_extrap.out

Any idea what's going wrong?

Example .bam and .bed files attached.
bams.zip

@idot
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idot commented Feb 22, 2022

I have the same issue. I use the preseq from bioconda which was built with --enable-hts .

wget https://depot.galaxyproject.org/singularity/preseq%3A3.1.2--h06ef8b0_1
singularity exec preseq:3.1.2--h06ef8b0_1 preseq  lc_extrap -pe -output 187530_T1.ccurve.txt  187530_T1.bam
ERROR:	max count before zero is less than min required count (4) duplicates removed

see also #49

(I also tried preseq:3.1.2--hfb6f838_0)

@z-spider
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need -B or --bam,then we will encounter this problem: #57
but, -B option is not what was causing the issue.

preseq try to merge the paired-end reads, but there are multiple alignments in the bam. -v option can show that.
use samtools to fileter multiple alignments can fix it.

All the best.

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