Unconfigured Ad

Collapse
X
 
  • Time
  • Show
Clear All
new posts
  • fchatonnet
    Member
    • Sep 2014
    • 30

    #1

    Tophat on Cygwin error

    Hello and thank you to welcome me in the community. I have been a intensive user of seqanswers threads to answer my many problems with bioinformatics, but this time, I have found no solution.
    I want to perform colorspace alignments using something else than LifeScope (the SOLiD dedicated out-of-date software). As I used tophat before for Illumina, I wanted to try a combination tophat2 / bowtie1 (bowtie2 does not handle colorspace). So after many problems, I finally got bowtie (v1.1.0) and tophat (2.0.12) running. Well bowtie runs, but not tophat, I get this error:

    $ more tophat_out/logs/tophat.log

    [2014-09-04 16:15:53] Beginning TopHat run (v2.0.12)
    -----------------------------------------------
    [2014-09-04 16:15:53] Checking for Bowtie
    Bowtie version: 1.1.0.0
    [2014-09-04 16:15:54] Checking for Samtools
    Samtools version: 0.1.19.0
    [2014-09-04 16:15:54] Checking for Bowtie index files (genome)..
    [2014-09-04 16:15:54] Checking for reference FASTA file
    [2014-09-04 16:15:54] Generating SAM header for mm10C
    [2014-09-04 16:15:54] Preparing reads
    left reads: min. length=50, max. length=50, 98 kept reads (2 discarded)
    [2014-09-04 16:15:54] Mapping left_kept_reads to genome mm10C with Bowtie
    [FAILED]
    Error running bowtie:
    Could not open single-ended aligned/unaligned-read file for writing: /dev/null

    This application has requested the Runtime to terminate it in an unusual way.
    Please contact the application's support team for more information.
    terminate called after throwing an instance of 'int'
    Last lign of run.log says:

    /usr/sbin/bam2fastx --all --color ./tophat_out/tmp/left_kept_reads.bam|/usr/sbin/bowtie -C --col-keepends -v 2 -k 20 -m 20 -S -p 1 --sam-nohead --max /dev/null mm10C -|/usr/sbin/fix_map_ordering --color --read-
    mismatches 2 --read-gap-length 2 --read-edit-dist 2 --read-realign-edit-dist 3 --index-outfile ./tophat_out/tmp/left_kept_reads.mapped.bam.index --bowtie1 --sam-header ./tophat_out/tmp/mm10C_genome.bwt.samheade
    r.sam - ./tophat_out/tmp/left_kept_reads.mapped.bam ./tophat_out/tmp/left_kept_reads_unmapped.bam
    I decomposed the actions, bam2fastx runs well, bowtie also as long as I change /dev/null to something else. I suspect a permission problem.
    I am under cygwin 2.850 64 bits, tried to move all my cygwin tree under an admin session and got the same problem. Any ideas?
    If not I will try to install another colorspace-able aligner (any suggestion?)

    Thanks a lot if you read the whole message and for your help (?!)
  • WhatsOEver
    Senior Member
    • Apr 2012
    • 215

    #2
    You just provide a folder and not a filename to the "max" parameter and that's what the program is complaining about

    [FAILED]
    Error running bowtie:
    Could not open single-ended aligned/unaligned-read file for writing: /dev/null
    /usr/sbin/bam2fastx --all --color ./tophat_out/tmp/left_kept_reads.bam|/usr/sbin/bowtie -C --col-keepends -v 2 -k 20 -m 20 -S -p 1 --sam-nohead --max /dev/null mm10C -|/usr/sbin/fix_map_ordering --color --read-
    mismatches 2 --read-gap-length 2 --read-edit-dist 2 --read-realign-edit-dist 3 --index-outfile ./tophat_out/tmp/left_kept_reads.mapped.bam.index --bowtie1 --sam-header ./tophat_out/tmp/mm10C_genome.bwt.samheade
    r.sam - ./tophat_out/tmp/left_kept_reads.mapped.bam ./tophat_out/tmp/left_kept_reads_unmapped.bam

    Comment

    • fchatonnet
      Member
      • Sep 2014
      • 30

      #3
      Except that --max is a bowtie option and I don't know how to specify it when I run tophat. As said in my previous most, if I specify --max with a file name when running the bowtie subroutine, everything goes well.

      Comment

      • WhatsOEver
        Senior Member
        • Apr 2012
        • 215

        #4
        ok, sry, didn't got that before. Please paste the command you used to run tophat then.

        Comment

        • fchatonnet
          Member
          • Sep 2014
          • 30

          #5
          Here is the tophat command line

          tophat -C -Q --bowtie1 --library-type fr-secondstrand mm10C toto.csfasta toto.QV.qual
          Then, as I said, I decomposed the last line of the run.log: bam2fastx works fine, used the output as input for bowtie, works also fine with --max toto_unmapped.fq as option. fix_map_ordering was also tested, but not directly with bowtie output.
          Thanks for your reply by the way.

          Comment

          • WhatsOEver
            Senior Member
            • Apr 2012
            • 215

            #6
            Looks like a cygwin related problem then.
            /dev/null should be accessible without special permissions.
            Only thing that comes into my mind is to provide full paths to everything and specify also an output dir with fullpath ("-o") for tophat files. Using --keep-tmp tmp files created during the tophat run won't be deleted. Within those you may also find additional information about what is happening.

            Comment

            Latest Articles

            Collapse

            • SEQadmin2
              Multiomic Liquid Biopsy: What DNA/RNA Panels Reveal That DNA Alone Can’t
              by SEQadmin2



              Tumors contain complex mixtures of cell populations that evolve in response to treatment and host-specific factors. Tissue biopsies provide only static snapshots of a tumor’s state. But what if we could follow that evolution over time using a minimally invasive technique?

              Liquid biopsies can detect tumor-derived material circulating in the bloodstream, providing a way to study treatment response, detect emerging resistance, and discover new biomarkers. Circulating
              ...
              10-14-2026, 09:55 AM
            • SEQadmin2
              New Genomics Technologies Take Aim at Long-Standing Limits
              by SEQadmin2


              Researchers using sequencing and genomics tools often have to make trade-offs. They can choose between speed or scale, short reads or long-range information, or targeted panels or a view of the whole transcriptome. New technologies that have been released this year are built to address those tough choices.

              We asked six companies the same four questions to learn about their latest products. The new technologies bring a lot to the table, including rethinking sequencing
              ...
              09-28-2026, 10:25 AM

            ad_right_rmr

            Collapse

            News

            Collapse

            Topics Statistics Last Post
            Started by SEQadmin2, Yesterday, 11:13 AM
            0 responses
            19 views
            0 reactions
            Last Post SEQadmin2  
            Started by SEQadmin2, Yesterday, 10:48 AM
            0 responses
            18 views
            0 reactions
            Last Post SEQadmin2  
            Started by SEQadmin2, 10-15-2026, 10:52 AM
            0 responses
            36 views
            0 reactions
            Last Post SEQadmin2  
            Started by SEQadmin2, 10-14-2026, 05:39 AM
            0 responses
            42 views
            0 reactions
            Last Post SEQadmin2  
            Working...