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  • jiangs
    Junior Member
    • Aug 2013
    • 2

    #1

    Tophat :OSError: [Errno 2] No such file or directory: '/tmp/segment.juncs

    Greetings,
    I'm new to tophat, here is my problem and I really need your help.
    My command is tophat2 -o NA06993 -G Ensemble.655.gtf -a 8 -m 0 -I 300000 -p 2 -g 20 --library-type fr-unstranded --fusion-read-mismatches 3 /rd/data/iGenomes/Homo_sapiens/UCSC/hg19/Sequence/Bowtie2Index/genome SRR032239.fastq

    The error information is :
    Searching for junctions via segment mapping
    Coverage-search algorithm is turned on, making this step very slow
    Please try running TopHat again with the option (--no-coverage-search) if this step takes too much time or memory.
    Traceback (most recent call last):
    File "/rd/tools_centos6.3/build/tophat-2.0.9/install/bin/tophat", line 4072, in <module>
    sys.exit(main())
    File "/rd/tools_centos6.3/build/tophat-2.0.9/install/bin/tophat", line 4038, in main
    user_supplied_deletions)
    File "/rd/tools_centos6.3/build/tophat-2.0.9/install/bin/tophat", line 3614, in spliced_alignment
    if os.path.getsize(juncs[0]) != 0:
    File "/usr/lib64/python2.6/genericpath.py", line 49, in getsize
    return os.stat(filename).st_size
    OSError: [Errno 2] No such file or directory: 'NA06993/tmp/segment.juncs'

    So how can I fix this?
  • genome1
    Member
    • Oct 2013
    • 12

    #2
    I am also facing similar problem while running tophat2.
    I am using tophat2 (v2.0.11) and Bowtie2 (v2.2.2).
    I am using default parameters while running tophat. My command line is:
    $ tophat -G /path/to/Homo_sapiens/Ensembl/GRCh37/Annotation/Genes/genes.gtf -o /path/to/out -p 8 /path/to/Homo_sapiens/Ensembl/GRCh37/Sequence/Bowtie2Index/genome /path/to/read_1.fastq.gz /path/to/read_2.fastq.gz

    The last few lines of the error message is:

    [2014-04-18 01:59:45] Resuming TopHat pipeline with unmapped reads
    Traceback (most recent call last):
    File "./tophat", line 4084, in <module>
    sys.exit(main())
    File "./tophat", line 4050, in main
    user_supplied_deletions)
    File "./tophat", line 3452, in spliced_alignment
    if not nonzeroFile(initial_reads[0]) and \
    File "./tophat", line 1159, in nonzeroFile
    samtools_view = subprocess.Popen(samtools_view_cmd, stdout=subprocess.PIPE)
    File "/usr/lib64/python2.6/subprocess.py", line 642, in __init__
    errread, errwrite)
    File "/usr/lib64/python2.6/subprocess.py", line 1234, in _execute_child
    raise child_exception
    OSError: [Errno 2] No such file or directory

    Can anyone help me to solve the problem.
    Thank you.

    Comment

    • GenoMax
      Senior Member
      • Feb 2008
      • 7142

      #3
      Is this a new install? If it is, have you tested it with the sample data to ensure everything is working properly.

      Comment

      • genome1
        Member
        • Oct 2013
        • 12

        #4
        The problem has been resolved.
        Now I have used tophat2 instead of tophat in my command line.

        Comment

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