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samtools-ampliconclip(1)      Bioinformatics tools	samtools-ampliconclip(1)

NAME
     samtools ampliconclip - clip reads using a BED file

SYNOPSIS
     samtools  ampliconclip  [-o out.file] [-f stat.file] [--soft-clip] [--hard-
     clip] [--both-ends]  [--strand]  [--clipped]  [--fail]  [--filter-len  INT]
     [--fail-len  INT]	[--unmap-len  INT]  [--no-excluded]  [--rejects-file re-
     jects.file]  [--original]	[--keep-tag]  [--tolerance]  [--no-PG]	[-u]  -b
     bed.file in.file

DESCRIPTION
     Clips the ends of read alignments if they intersect with regions defined in
     a	BED  file.   While  this  tool	was originally written for clipping read
     alignment positions which correspond to amplicon primer  locations  it  can
     also be used in other contexts.

     BED  file	entries  used  are  chrom, chromStart, chromEnd and, optionally,
     strand.  Standard BED file format must be used, so if strand is needed then
     the name and score fields must also be present  (even  though  ampliconclip
     does not read them).  There is a default tolerance of 5 bases when matching
     chromStart and chromEnd to alignments.

     By  default  the  reads  are soft clipped and clip is only done from the 5'
     end.

     Some things to be aware of.  While ordering is not significant, adjustments
     to the left most mapping position (POS) will mean	that  coordinate  sorted
     files  will  need resorting.  In such cases the sorting order in the header
     is set to unknown. Clipping of reads results in template length (TLEN)  be-
     ing  incorrect.  This can be corrected by samtools fixmates.  Any MD and NM
     aux tags will also be incorrect, which can be fixed by samtools calmd.   By
     default  MD  and NM tags are removed though if the output is in CRAM format
     these tags will be automatically regenerated.

OPTIONS
     -b FILE	BED file of regions (e.g. amplicon primers) to be removed.

     -o FILE	Output file name (defaults to stdout).

     -f FILE	File to write stats to (defaults to stderr).

     -u 	Output uncompressed SAM, BAM or CRAM.

     --soft-clip
		Soft clip reads (default).

     --hard-clip
		Hard clip reads.

     --both-ends
		Clip at both the 5' and the 3' ends where regions  match.   When
		using this option the --strand option is ignored.

     --strand	Use  strand entry from the BED file to clip on the matching for-
		ward or reverse alignment.

     --clipped	Only output clipped reads.  Filter all others.

     --fail	Mark unclipped reads as QC fail.

     --filter-len INT
		Filter out reads of INT size or  shorter.   In	this  case  soft
		clips are not counted toward read length.  An INT of 0 will fil-
		ter out reads with no matching bases.

     --fail-len INT
		As --filter-len but mark as QC fail rather then filter out.

     --unmap-len INT
		As  --filter-len but mark as unmapped. Default is 0 (no matching
		reads).  -1 will disable.

     --no-excluded
		Filter out any reads that are marked as QCFAIL or are  unmapped.
		This  works  on  the  state  of  the reads before clipping takes
		place.

     --rejects-file FILE
		Write any filtered reads out to a file.

     --primer-counts FILE
		File to write with read counts per bed entry (bedgraph format).

     --original
		Add an OA tag with the original data for clipped files.

     --keep-tag
		In clipped reads, keep the possibly invalid NM and MD tags.   By
		default these tags are deleted.

     --tolerance INT
		The  amount of latitude given in matching regions to alignments.
		Default 5 bases.

     --no-PG	Do not at a PG line to the header.

AUTHOR
     Written by Andrew Whitwham and Rob Davies, both from the Sanger Institute.

SEE ALSO
     samtools(1), samtools-sort(1), samtools-fixmate(1), samtools-calmd(1)

     Samtools website: <http://www.htslib.org/>

samtools-1.23.1 		  18 March 2026 	samtools-ampliconclip(1)

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