vsearch

Versatile open-source tool for microbiome analysis

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NAME

vsearch \-\-fastq_join — join paired-end reads into one sequence with a gap

SYNOPSIS

vsearch \-\-fastq_join fastqfile \-\-reverse fastqfile (\-\-fastaout | \-\-fastqout) filename [options]

DESCRIPTION

The vsearch command --fastq_join joins paired-end reads into a single sequence. Forward reads are given as the argument to --fastq_join, and the reverse reads are specified with --reverse. The resulting sequence consists of the forward read, a padding sequence, and the reverse complement of the reverse read. To illustrate:

>R1            >R2                  >R1
AAACCC    +    GGGAAA    -->   AAACCCNNNNNNNNTTTCCC
                               (R1) (padding) (revcomp R2)

The padding sequence defaults to NNNNNNNN (eight N’s) and can be changed with --join_padgap. The corresponding quality string defaults to IIIIIIII (quality score 40, error probability 0.0001) and can be changed with --join_padgapq. Joined sequences are written to --fastaout and/or --fastqout. See vsearch-fasta(5) and vsearch-fastq(5) for more information on these formats.

Note that reads are joined, not merged. To merge overlapping paired-end reads, see vsearch-fastq_mergepairs(1).

OPTIONS

mandatory options

--reverse filename
Specify the FASTQ file containing the reverse reads.

At least one of --fastaout or --fastqout must also be specified.

core options

--fastaout filename
Write the joined sequences to filename, in fasta format (see vsearch-fasta(5)). Quality scores are not written.

--fastqout filename
Write the joined sequences to filename, in fastq format (see vsearch-fastq(5)).

--join_padgap string
Use string as the sequence padding inserted between the forward and reverse reads. The default is NNNNNNNN (eight N’s). Accepts an empty string, or any other combination of ASCII characters.

--join_padgapq string
Use string as the quality padding inserted between the forward and reverse reads. The default is a string of I’s of the same length as the sequence padding string (see --join_padgap). The letter I corresponds to a base quality score of 40, indicating a very high quality base with error probability of 0.0001 (see vsearch-expected_error(7)). Accepts an empty string, or any other combination of ASCII characters.

secondary options

--bzip2_decompress
Specify that the input pipe is streaming data compressed using Huffman coding. See bzip2(1) for more details. This option is not needed when reading from a regular file compressed with bzip2.

--fasta_width positive integer
Set the maximal width of sequences when writing fasta files. Longer sequences are folded and written on several lines. Default width is 80 nucleotides. Set to zero (0) to suppress folding.

--fastq_ascii 33|64
Specify the offset used as the basis for the fastq quality score when reading fastq files. For example, an offset of 33 means that a quality value of 41 is represented by the 74th ASCII symbol (33 + 41 = 74), which is ‘J’. See ascii(7) for a view of the ASCII character set. The offset value is either 33 or 64, default is 33.

--gzip_decompress
Specify that the input pipe is streaming data compressed using Lempel-Ziv coding. See gzip(1) for more details. This option is not needed when reading from a regular file compressed with gzip.

--label_suffix string
Add the suffix string to sequence headers when writing fasta or fastq files. For example, with --label_suffix ";status=healthy", sequence header ‘>seq1’ becomes ‘>seq1;status=healthy’.

--lengthout
Add a sequence length annotation (;length=integer) to each sequence header when writing fasta or fastq files.

--log filename
Write messages to filename. Messages include program version, start and finish times, elapsed time, amount of memory available, maximum amount of memory consumed, number of cores and command line options, and if need be, command-specific informational messages, warnings, and errors.

--no_progress
Suppress the gradually increasing progress indicator normally written to the standard error stderr(3).

--quiet
Suppress messages to the standard output stdout(3) and standard error stderr(3), except for warnings and error messages.

--relabel string
Replace sequence headers with the prefix string and a ticker (1, 2, 3, etc.). For example, with --relabel "cluster:", the first sequence header becomes ‘>cluster:1’, the second sequence header becomes ‘>cluster:2’, and so on. To retain annotations, use their corresponding options (--lengthout, --eeout, and --sizeout). Use --relabel_keep to also retain old sequence identifiers.

--relabel_keep
Retain old sequence identifiers by including them at the end of the new headers, after a space.

--relabel_md5
Replace each sequence header with the MD5 digest derived from the sequence itself. The sequence is converted to upper case, and each ‘U’ is replaced with a ‘T’ before computation of the digest. The MD5 digest is a 128-bit value (16 bytes), represented using a string of 32 ASCII characters. Each pair of characters encodes an hexadecimal value, ranging from x00 to xff. See md5(3) for more details, and --relabel_sha1 for an alternative hashing algorithm. To retain annotations, use their corresponding options (--lengthout, --eeout, and --sizeout). Use --relabel_keep to also retain old sequence identifiers.

--relabel_self
Replace each sequence header with the sequence itself. To retain annotations, use their corresponding options (--lengthout, --eeout, and --sizeout). Use --relabel_keep to also retain old sequence identifiers.

--relabel_sha1
Replace each sequence header with the SHA1 digest derived from the sequence itself. The sequence is converted to upper case, and each ‘U’ is replaced with a ‘T’ before computation of the digest. The SHA1 digest is a 160-bit value (20 bytes), represented using a string of 40 ASCII characters. Each pair of characters encodes an hexadecimal value, ranging from x00 to xff. See sha1(3) for more details, and --relabel_md5 for an alternative hashing algorithm. To retain annotations, use their corresponding options (--lengthout, --eeout, and --sizeout). Use --relabel_keep to also retain old sequence identifiers.

--sizein
Use the abundance annotations present in sequence headers when reading fasta or fastq file. Search for the pattern [>@;]size=integer[;]. Entries without abundance annotations are silently assumed to be of size=1.

--sizeout
Add abundance annotations to sequence headers when writing fasta or fastq files. Add the pattern ;size=integer. If option --sizein is not used, abundance values are set to 1 for all entries. If --sizein is used, existing abundance annotations are simply reported to output files.

--xee
Strip expected error (ee) annotations from sequence headers when writing fasta or fastq files. Search for the pattern [>@;]ee=float[;]. Expected error annotations are added by the synonymous options --fastq_eeout and --eeout described in vsearch-fastx_filter(1).

--xlength
Strip sequence length annotations from sequence headers when writing fasta or fastq files. Search for the pattern [>@;]length=integer[;]. Sequence length annotations are added by the --lengthout option.

--xsize
Strip abundance annotations from sequence headers when writing fasta or fastq files. Search for the pattern [>@;]size=integer[;]. Abundance annotations are added by the --sizeout option.

ignored options

--fastq_qmax positive integer
Option is ignored and has no effect.

--fastq_qmin positive integer
Option is ignored and has no effect.

--threads positive non-null integer
Command is not multithreaded, option has no effect.

EXAMPLES

Join forward reads in R1.fastq with reverse reads in R2.fastq, and write the joined sequences to joined.fastq:

vsearch \
    --fastq_join R1.fastq \
    --reverse R2.fastq \
    --fastqout joined.fastq

Use a custom padding sequence of twelve N’s and a matching quality string of twelve I’s:

vsearch \
    --fastq_join R1.fastq \
    --reverse R2.fastq \
    --join_padgap NNNNNNNNNNNN \
    --join_padgapq IIIIIIIIIIII \
    --fastqout joined.fastq

SEE ALSO

vsearch-fastq_mergepairs(1) for merging overlapping paired-end reads. vsearch-fasta(5), vsearch-fastq(5), vsearch-expected_error(7)

CITATION

Rognes T, Flouri T, Nichols B, Quince C, Mahé F. (2016) VSEARCH: a versatile open source tool for metagenomics. PeerJ 4:e2584 doi: 10.7717/peerj.2584

REPORTING BUGS

Submit suggestions and bug-reports at https://github.com/torognes/vsearch/issues, send a pull request on https://github.com/torognes/vsearch, or compose a friendly or curmudgeont e-mail to Torbjørn Rognes (torognes@ifi.uio.no).

AVAILABILITY

Source code and binaries are available at https://github.com/torognes/vsearch.

COPYRIGHT

Copyright (C) 2014-2026, Torbjørn Rognes, Frédéric Mahé and Tomás Flouri

All rights reserved.

Contact: Torbjørn Rognes torognes@ifi.uio.no, Department of Informatics, University of Oslo, PO Box 1080 Blindern, NO-0316 Oslo, Norway

This software is dual-licensed and available under a choice of one of two licenses, either under the terms of the GNU General Public License version 3 or the BSD 2-Clause License.

GNU General Public License version 3

This program is free software: you can redistribute it and/or modify it under the terms of the GNU General Public License as published by the Free Software Foundation, either version 3 of the License, or (at your option) any later version.

This program is distributed in the hope that it will be useful, but WITHOUT ANY WARRANTY; without even the implied warranty of MERCHANTABILITY or FITNESS FOR A PARTICULAR PURPOSE. See the GNU General Public License for more details.

You should have received a copy of the GNU General Public License along with this program. If not, see http://www.gnu.org/licenses/.

The BSD 2-Clause License

Redistribution and use in source and binary forms, with or without modification, are permitted provided that the following conditions are met:

  1. Redistributions of source code must retain the above copyright notice, this list of conditions and the following disclaimer.

  2. Redistributions in binary form must reproduce the above copyright notice, this list of conditions and the following disclaimer in the documentation and/or other materials provided with the distribution.

THIS SOFTWARE IS PROVIDED BY THE COPYRIGHT HOLDERS AND CONTRIBUTORS “AS IS” AND ANY EXPRESS OR IMPLIED WARRANTIES, INCLUDING, BUT NOT LIMITED TO, THE IMPLIED WARRANTIES OF MERCHANTABILITY AND FITNESS FOR A PARTICULAR PURPOSE ARE DISCLAIMED. IN NO EVENT SHALL THE COPYRIGHT HOLDER OR CONTRIBUTORS BE LIABLE FOR ANY DIRECT, INDIRECT, INCIDENTAL, SPECIAL, EXEMPLARY, OR CONSEQUENTIAL DAMAGES (INCLUDING, BUT NOT LIMITED TO, PROCUREMENT OF SUBSTITUTE GOODS OR SERVICES; LOSS OF USE, DATA, OR PROFITS; OR BUSINESS INTERRUPTION) HOWEVER CAUSED AND ON ANY THEORY OF LIABILITY, WHETHER IN CONTRACT, STRICT LIABILITY, OR TORT (INCLUDING NEGLIGENCE OR OTHERWISE) ARISING IN ANY WAY OUT OF THE USE OF THIS SOFTWARE, EVEN IF ADVISED OF THE POSSIBILITY OF SUCH DAMAGE.

ACKNOWLEDGMENTS

We would like to thank the authors of the following projects for making their source code available: