The Institute for Systems Biology RepeatModeler Download

RepeatModeler

RepeatModeler is a de novo transposable element (TE) family identification and modeling package. At the heart of RepeatModeler are three de-novo repeat finding programs ( RECON, RepeatScout and LtrHarvest/Ltr_retriever ) which employ complementary computational methods for identifying repeat element boundaries and family relationships from sequence data.

RepeatModeler assists in automating the runs of the various algorithms given a genomic database, clustering redundant results, refining and classifying the families and producing a high quality library of TE families suitable for use with RepeatMasker and ultimately for submission to the Dfam database ( http://dfam.org ).

Authors

RepeatModeler: Robert Hubley, Arian Smit - Institute for Systems Biology

LTR Pipeline Extensions: Jullien M. Flynn - Cornell University

Installation

There are two supported paths to installing RepeatModeler on a UNIX-based server. RepeatModeler may be installed from source as described in the Source Distribution Installation instructions below, or using one of our Dfam-TETools container images ( Docker or Singularity ). The containers include RepeatModeler, it’s prerequisites and additional TE analysis tools/utilities used by Dfam. Information on the Dfam-TETools container may be found here: https://github.com/Dfam-consortium/TETools

Source Distribution Installation

Prerequisites

Perl Available at http://www.perl.org/get.html. Developed and tested with version 5.8.8.

RepeatMasker & Libraries Developed and tested with open-4.1.4. The program is available at http://www.repeatmasker.org/RMDownload.html and is distributed with Dfam - an open database of transposable element families.

RECON - De Novo Repeat Finder, Bao Z. and Eddy S.R. Developed and tested with our patched version of RECON ( 1.08 ). The 1.08 version fixes problems with running RECON on 64 bit machines and supplies a workaround to a division by zero bug along with some buffer overrun fixes. The program is available at: http://www.repeatmasker.org/RepeatModeler/RECON-1.08.tar.gz. The original version is available at http://eddylab.org/software/recon/.

RepeatScout - De Novo Repeat Finder, Price A.L., Jones N.C. and Pevzner P.A. Developed and tested with our multiple sequence version of RepeatScout ( 1.0.6 ). This version is available at http://www.repeatmasker.org/RepeatScout-1.0.6.tar.gz

TRF - Tandem Repeat Finder, G. Benson et al. You can obtain a free copy at http://tandem.bu.edu/trf/trf.html RepeatModeler requires version 4.0.9 or higher.

RMBlast - A modified version of NCBI Blast for use with RepeatMasker and RepeatModeler. Precompiled binaries and source can be found at http://www.repeatmasker.org/RMBlast.html We highly recommend using 2.13.0 or higher.

UCSC genome browser command-line utilities - Some tools included with RepeatModeler work with files in the ‘twobit’ file format using these programs: twoBitToFa, faToTwoBit, and twoBitInfo. Precompiled binaries and source for these programs can be found at http://hgdownload.soe.ucsc.edu/downloads.html#utilities_downloads.

Optional. Required for running LTR structural search pipeline:

LtrHarvest - The LtrHarvest program is part of the GenomeTools suite. We have developed this release of RepeatModeler on GenomeTools version 1.5.9 available for download from here: http://genometools.org/pub/ NOTE: use the “make threads=yes” build options to enable multi-threaded runs.

Ltr_retriever - A LTR discovery post-processing and filtering tool. We recommend using version 2.6 or higher from here: https://github.com/oushujun/LTR_retriever/releases

MAFFT - A multiple sequence alignment program. We developed and tested RepeatModeler using mafft version 7.407. Please use this verison or higher from here: https://mafft.cbrc.jp/alignment/software/

CD-HIT - A sequence clustering package. We developed and tested RepeatModeler using version 4.8.1. Please use this version or higher from: http://weizhongli-lab.org/cd-hit/

Ninja - A tool for large-scale neighbor-joining phylogeny inference and clustering. We developed and tested RepeatModeler using Ninja version “0.95-cluster_only”. Please obtain a copy from: https://github.com/TravisWheelerLab/NINJA/releases/tag/0.95-cluster_only

Installation

  1. Obtain the source distribution

    • Github : https://github.com/Dfam-consortium/RepeatModeler Available by cloning the master branch of the RepeatModeler repository ( latest released version ) or by downloading a release from the repository “releases” tab. or
    • RepeatMasker Website : http://www.repeatmasker.org/RepeatModeler
  2. Uncompress and expand the distribution archive:

    tar -zxvf RepeatModeler-#.#.tar.gz
  3. Configure for your site:

    Automatic:

    • Run the “configure” script interactively with prompts for each setting:

        perl ./configure
    • Run the “configure” script with supplied paramters:

        perl ./configure -rscout_dir .. -recon_dir ..

    By Hand:

    • Edit the configuration file “RepModelConfig.pm”

    Dynamically:

    • Use the “configuration overrides” command line options with the RepeatModeler programs. e.g:

        ./RepeatModeler -rscout_dir .. -recon_dir .. 

Example Run

In this example we first downloaded elephant sequences from Genbank ( approx 11MB ) into a file called elephant.fa.

  1. Create a Database for RepeatModeler

    RepeatModeler uses a NCBI BLASTDB as input to the repeat modeling pipeline. A utility is provided to assist the user in creating a single database from several types of input structures.

      <RepeatModelerPath>/BuildDatabase -name elephant elephant.fa

    Run “BuildDatabase” without any options in order to see the full documentation on this utility. There are several options which make it easier to import multiple sequence files into one database.

    TIP: It is a good idea to place your datafiles and run this program suite from a local disk rather than over NFS. This will greatly improve runtime as the filesystem access is considerable

  2. Run RepeatModeler

    RepeatModeler runs several compute intensive programs on the input sequence. For best results run this on a single machine with a moderate amount of memory > 32GB and multiple processors.
    Our setup is Xeon(R) CPU E5-2680 v4 @ 2.40GHz - 28 cores, 128GB RAM. To specify a run using 20 threads (at most), and including the new LTR discovery pipeline:

       nohup <RepeatModelerPath>/RepeatModeler -database elephant 
       -threads 20 -LTRStruct >& run.out &

    The nohup (or screen) is used on our machines when running long jobs. The log output is saved to a file and the process is backgrounded. For typical runtimes ( can be > 2-3 days with this configuration on a well assembled mammalian genome ) see the run statistics section of this file. It is important to save the log output for later usage.
    It contains the random number generator seed so that the sampling process may be reproduced if necessary. In addition the log file contains details about the progress of the run for later assesment of peformance or debuging problems.

  3. Interpret the results

    RepeatModeler produces a voluminous amount of temporary files stored in a directory created at runtime named like:

          RM_<PID>.<DATE> ie. "RM_5098.MonMar141305172005" 

    and remains after each run for debugging purposes or for the purpose of resuming runs if a failure occures. At the succesful completion of a run, three files are generated:

          <database_name>-families.fa  : Consensus sequences
          <database_name>-families.stk : Seed alignments
          <database_name>-rmod.log     : A summarized log of the run

    The seed alignment file is in a Dfam compatible Stockholm format and may be uploaded to the Dfam database by submiting the data to help@dfam.org or by going to dfam.org/login and creating an upload account.

    The fasta format is useful for running quick custom library searches using RepeatMasker. Ie.:

          <RepeatMaskerPath>/RepeatMasker -lib <database_name>-families.fa mySequence.fa 

    Other files produced in the working directory include:

       RM_<PID>.<DATE>/
          consensi.fa
          families.stk
          round-1/
               sampleDB-#.fa       : The genomic sample used in this round
               sampleDB-#.fa.lfreq : The RepeatScout lmer table
               sampleDB-#.fa.rscons: The RepeatScout generated consensi
               sampleDB-#.fa.rscons.filtered : The simple repeat/low 
                                               complexity filtered
                                               version of *.rscons
               consensi.fa         : The final consensi db for this round
               family-#-cons.html  : A visualization of the model
                                     refinement process.  This can be opened
                                     in web browsers that support zooming.
                                     ( such as firefox ).
                                     This is used to track down problems 
                                     with the Refiner.pl
               index.html          : A HTML index to all the family-#-cons.html
                                     files.
          round-2/
               sampleDB-#.fa       : The genomic sample used in this round
               msps.out            : The output of the sample all-vs-all 
                                     comparison
               summary/            : The RECON output directory
                    eles           : The RECON family output
               consensi.fa         : Same as above
               family-#-cons.html  : Same as above
               index.html          : Same as above
          round-3/
               Same as round-2
           ..
          round-n/

Recover from a failure

If for some reason RepeatModeler fails, you may restart an analysis starting from the last round it was working on. The -recoverDir [ResultDir] option allows you to specify a diretory ( i.e RM_./ ) where a previous run of RepeatModeler was working and it will automatically determine how to continue the analysis.

Caveats

Please see the RELEASE-NOTES file for more details.

RepeatModeler Statistics

Benchmarks and statistics for runs of RepeatModeler on several sample genomes.

RepeatModeler 2.0 ( RECON + RepeatScout + LTRStruct ):

Genome Genome DB Size (bp) Runtime (hh:mm)* Models Built
D. melanogaster 164 Mbp 12:56 734
D. rerio 1.4 Gbp 40:36 3851
O. sativa 375 Mbp 37:23 2648

Previous Versions

RepeatModeler 1.0.3 ( RECON + RepeatScout ):

            Genome DB    Sample***   Run Time*   Models   Models       % Sample
Genome      Size (bp)    Size (bp)   (hh:mm)     Built    Classified   Masked**
----------  -----------  ----------  ----------  -------  -----------  --------
Marmoset    3.0 Bbp      228 Mbp      55:37        582      564         36.4    
Zebrafinch  1.2 Bbp      222 Mbp      66:29        178       75          8.6    


  *  Analysis run on a 4 processor P4, 2.4Ghz, 3GB RAM, machine
     running Red Hat Linux.

  ** Includes simple repeats and low complexity DNA. Results
     obtained with RepeatMasker open-3.2.5, WUBlast and
     the -lib option.

  *** Sample size does not include 40 Mbp used in the RepeatScout analysis.  
      This 40 Mbp is randomly chosen and may overlap 0-100% of the 
      sample used in the RECON analysis.


RepeatModeler 1.0.2 ( RECON + RepeatScout ):

            Genome DB    Sample***   Run Time*   Models   Models       % Sample
Genome      Size (bp)    Size (bp)   (hh:mm)     Built    Classified   Masked**
----------  -----------  ----------  ----------  -------  -----------  --------
Human HG18     3.1 Bbp      238 Mbp   46:36        614      611         35.66
Platypus       1.9 Bbp      220 Mbp   76:02        540      457         
Zebrafinch     1.3 Bbp      220 Mbp   63:57        233      104          9.41
Sea Urchin     867 Mbp      220 Mbp   40:03       1830      360         33.85
diatom       32,930,227  32,930,227    4:41        128       35          2.86
Rabbit       11,770,949  11,770,949    3:14         83       72         31.30

  *  Analysis run on a 4 processor P4, 2.4Ghz, 3GB RAM, machine
     running Red Hat Linux.

  ** Includes simple repeats and low complexity DNA. Results
     obtained with RepeatMasker open-3.1.9, WUBlast and
     the -lib option.

  *** Sample size does not include 40 Mbp used in the RepeatScout analysis.  
      This 40 Mbp is randomly chosen and may overlap 0-100% of the 
      sample used in the RECON analysis.



RepeatModeler 1.0.0 ( RECON ):

            Genome DB    Sample      Run Time*   Models   Models       % Sample
Genome      Size (bp)    Size (bp)   (hh:mm)     Built    Classified   Masked**
----------  -----------  ----------  ----------  -------  -----------  --------
Opossum     3.5 Billion  319 Mbp     140:52      1137     467          52.55
Armadillo   47,332,136   47,332,136    6:20      121      92           36.07
Platypus    14,768,992   14,768,992    3:46      18       13           40.69
Rabbit      11,770,949   11,770,949    2:17      20       16           28.67
Elephant    11,550,090   11,550,090    1:21      34       28           37.08

  *  Analysis run on a 4 processor P4, 2.4Ghz, 3GB RAM, machine
     running Red Hat Linux.
  ** Includes simple repeats and low complexity DNA. Results
     obtained with RepeatMasker open-3.0.9, WUBlast and
     the -lib option.

Credits

Arnie Kas for the work done on the original MultAln.pm.

Andy Siegel for statistics consultations.

Thanks so much to Warren Gish for his invaluable assistance and consultation on his ABBlast program suite.

Alkes Price and Pavel Pevzner for assistance with RepeatScout and hosting my multi-sequence version of RepeatScout.

Shujun Ou, and Ning Jiang for discussions and assistance with using LTR_retreiver.

This work was supported by the NIH ( R44 HG02244-02), ( RO1 HG002939 ), ( U24 HG010136 ), and the Institute for Systems Biology.

License

This work is licensed under the Open Source License v2.1.
To view a copy of this license, visit http://www.opensource.org/licenses/osl-2.1.php or see the LICENSE file contained in this distribution.


Institute for Systems Biology
This server is made possible by funding from the National Human Genome Research Institute (NIGRI grant # RO1 HG002939).