NAME
    copyrighter - Correct trait bias in microbial profiles

SYNOPSIS
      copyrighter -i otu_table.qiime -o otu_table_copyrighted.generic

DESCRIPTION
    The genome of Bacteria and Archaea often contains several copies of the
    16S rRNA gene. This can lead to significant biases when estimating the
    composition of microbial communities using 16S rRNA amplicons or
    microarrays or their total abundance using 16S rRNA quantitative PCR,
    since species with a large number of copies will contribute
    disproportionally more 16S amplicons than species with a unique copy.
    Fortunately, it is possible to infer the copy number of unsequenced
    microbial species, based on that of close relatives that have been fully
    sequenced. Using this information, CopyRigher corrects microbial
    relative abundance by applying a weight proportional to the inverse of
    the estimated copy number to each species.

    In metagenomic surveys, a similar problem arises due to genome length
    variations between species, and can be corrected by CopyRighter as well.

    In all cases, a community file is used as input (-i option) and a
    corrected community file with trait-corrected (16S rRNA gene copy number
    or genome length) relative abundances is generated (-o option). Total
    abundance can optionally be provided (-t option), corrected and combined
    with relative abundance estimates to get the absolute abundance of each
    species. Also the average trait value in each community is reported on
    standard output.

    We are grateful to the Genomics Virtual Lab <https://genome.edu.au/> for
    providing a public Galaxy webserver in which users can run CopyRighter
    in a graphical environment: <http://galaxy-qld.genome.edu.au>.

REQUIRED ARGUMENTS
    -i <input>
        Input community file obtained from 16S rRNA microarray, 16S rRNA
        amplicon sequencing or metagenomic sequencing, in biom, QIIME, GAAS,
        Unifrac, or generic (tabular site-by-species) format. The file must
        contain read counts (not percentages) and taxa must have UNALTERED
        taxonomic assignments. Here is an example of Greengenes 2012/10
        taxonomic string (note the whitespace after each semicolon):

          k__Bacteria; p__Proteobacteria; c__Alphaproteobacteria; o__Rhodospirillales; f__Rhodospirillaceae; g__Telmatospirillum; s__siberiense

        See also the <data> parameter to specify your own database of trait
        values.

OPTIONAL ARGUMENTS
    -d <data>
        Provide the file of trait estimates to use for correction. Data
        files of 16S rRNA gene copy number and genome length (based on IMG
        4.0 genomes mapped onto the Oct 2012 Greengenes taxonomy) are
        distributed with CopyRighter. In case you want to use an alternative
        data file, be aware that it should be tab-delimited and have two
        columns, an ID or taxonomic string (col 1), and trait estimate (col
        2), as illustrated in this example:

          # ID  16S rRNA count
          4     1.51098055313977
          7     1.51812891020048
          ...
          24084 3.41268502385832

          # taxstring   16S rRNA count
          k__Archaea; p__; c__; o__; f__; g__; s__      1.57262
          k__Archaea; p__Crenarchaeota; [...] g__Cenarchaeum; s__symbiosum      1.00000
          ...
          k__Bacteria; p__Actinobacteria; [...] g__Actinomyces; s__europaeus    1.19211

        Extra columns are ignored, as well as empty lines and comment lines
        (starting with #). Note that the header line can define the name of
        the weight used. Also, the file can contain trait values both at the
        ID and taxstring level.

        This argument is optional. When omitted, CopyRighter will look for
        the data file location stored in the "COPYRIGHTER_DB" environment
        variable. Feel free to make this variable point to your preferred
        data file.

    -l <lookup>
        What to match when looking up the trait value of a taxon: 'desc',
        use taxonomic description, or 'id', use OTU ID (if recorded in your
        input community file). The script bc_use_repr_id of Bio::Community
        can help in replacing arbitrary OTU IDs by their corresponding
        Greengenes ID. Default: desc

    -o <output>
        Output path for the corrected community files (in same format as
        input), with relative abundance expressed in percent. Default:
        out_copyrighted.txt

    -t <total>
        File containing the total microbial abundance to be corrected by the
        average trait value, e.g. 16S rRNA quantitative PCR numbers to be
        corrected by the average 16S rRNA copy number in each community.
        This file should be tab-delimited and contain two columns: community
        name, and total abundance. Using this option will produce two
        additional output files, one containing the corrected total
        microbial abundance, and other the absolute abundance of each taxon
        in the <input> (in the same format as <input>): assuming an <output>
        called 'out_copyrighted.txt', these files will be named,
        respectively, 'out_copyrighted_total.tsv' and
        'out_copyrighted_combined.txt'.

    -v  Verbose mode. Display trait value assignments. You should probably
        use this option and make sure that your taxa are processed as
        intended.

HELP & FEEDBACK
  Mailing list
    New releases of CopyRighter, usage help and suggestions are discussed on
    this mailing list: <https://groups.google.com/d/forum/copyrighter>

  Bugs
    All complex software has bugs lurking in it, and this program is no
    exception. If you find a bug, please report it on the bug tracker:
    <http://github.com/fangly/AmpliCopyrighter/issues>

AUTHOR
    Florent Angly <florent.angly@gmail.com>

VERSION
    This document refers to copyrighter version 0.46

COPYRIGHT
    Copyright 2012-2014 Florent ANGLY <florent.angly@gmail.com>

    CopyRighter 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. CopyRighter 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 CopyRighter. If not,
    see <http://www.gnu.org/licenses/>.