Manual page from samtools-1.19
released on 12 December 2023


samtools split – splits a file by read group.


samtools split [options] merged.sam|merged.bam|merged.cram


Splits a file by read group, or a specified tag, producing one or more output files matching a common prefix (by default based on the input filename).

Unless the -d option is used, the file will be split according to the @RG tags listed in the header. Records without an RG tag or with an RG tag undefined in the header will cause the program to exit with an error unless the -u option is used.

RG values defined in the header but with no records will produce an output file only containing a header.

If the -d TAG option is used, the file will be split on the value in the given aux tag. Note that only string tags (type Z) are currently supported. Unless the -u option is used, the program will exit with an error if it finds a record without the given tag.

Note that attempting to split on a tag with high cardinality may result in the creation of a large number of output files. To prevent this, the -M option can be used to set a limit on the number of splits made.

Using -d RG behaves in a similar way to the default (without -d), opening an output file for each @RG line in the header. However, unlike the default, new output files will be opened for any RG tags found in the alignment records irrespective of if they have a matching header @RG line.

The -u option may be used to specify the output filename for any records with a missing or unrecognised tag. This option will always write out a file even if there are no records.

Output format defaults to BAM. For SAM or CRAM then either set the format with --output-fmt or use -f to set the file extension e.g. -f %*_%#.sam.


-u FILE1

Put reads with no tag or an unrecognised tag into FILE1

-h FILE2

Use the header from FILE2 when writing the file given in the -u option. This header completely replaces the one from the input file. It must be compatible with the input file header, which means it must have the same number of references listed in the @SQ lines and the references must be in the same order and have the same lengths.


Output filename format string (see below) ["%*_%#.%."]

-d TAG

Split reads by TAG value into distinct files. Only the TAG key must be supplied with the option. The value of the TAG has to be a string (i.e. key:Z:value )

-M,--max-split NUM

Limit the number of files created by the -d option to NUM (default 100). This prevents accidents where trying to split on a tag with high cardinality could result in the creation of a very large number of output files. Once the file limit is reached, any tag values not already seen will be treated as unmatched and the program will exit with an error unless the -u option is in use.

If desired, the limit can be removed using -M -1, although in practice the number of outputs will still be restricted by system limits on the number of files that can be open at once.

If splitting by read group, and the read group count in the header is higher than the requested limit then the limit will be raised to match.


Verbose output


Do not add a @PG line to the header of the output file.

Format string expansions:

%#index (of @RG in the header, or count of TAG values seen so far)
%!@RG ID or TAG value
%.output format filename extension
-@, --threads INT

Number of input/output compression threads to use in addition to main thread [0].


Written by Martin Pollard from the Sanger Institute.


samtools (1), samtools-addreplacerg (1)

Samtools website: <>