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  • Error when calling fusion junctions with MapSplice

    We want to compare the results of different tools that call splice junctions using RNA-seq data.

    Now, I try to call fusion junctions with MapSplice (1.15.2).

    My input reads are 100nt in length and in fastq format.

    Example:

    Code:
    @xxxyyyzzz
    AGATGCACTACCACCCAGCCTATCAGGACAGGCCAAGCCTGAGGACAGTGACTGTCACAGAAAAATAGAAACTTGTGGTTCCAGGAAATCCGAGAGGTCT
    +
    IIHIHIHHIHHIHHGHHFGIGHGGGGIIIHHNKIJKIJELHCJFFGLIMLHLGLIAHBIEGFFDJQDAE>HIJHOIMLHAHGGHMPGHIFEJMDVJBEOM
    I'm calling Mapsplice as follows (basically copy and pasted from the MapSpliceManual, just added the --fusion parameter):

    Code:
    python mapsplice_segments.py -u reads.fq -c [somewhere]/genome/hg19/singleChroms/ -B [somewhere]/genome/bowtie/hg19 -Q fq -X 5 -L 25 --fusion
    MapSplice quits with an error:

    Code:
    [Thu Oct 25 11:24:45 2012] Checking for chromosomes files or directory
    [Thu Oct 25 11:24:45 2012] Checking for chromosomes files or directory passed
    [Thu Oct 25 11:24:45 2012] Checking for Bowtie index files
    [Thu Oct 25 11:24:45 2012] check reads format
    [Thu Oct 25 11:24:46 2012] merge paired end reads remove short
    [Thu Oct 25 11:24:56 2012] Mapping reads against hg19 with Bowtie
    [Thu Oct 25 11:25:13 2012] Converting bowtie mapped to SAM format
    [Thu Oct 25 11:25:37 2012] divide reads
    [Thu Oct 25 11:25:56 2012] Mapping reads against hg19 with Bowtie
    [Thu Oct 25 11:26:50 2012] sort segmentbwt
    [Thu Oct 25 11:27:15 2012] reads all chromo sizes
    [Thu Oct 25 11:27:20 2012] mapsplice_search
    [Thu Oct 25 11:30:06 2012] Aligning spliced reads
    [Thu Oct 25 11:46:59 2012] mapsplice_report
    error: open bwtmap file mapsplice_out/tmp/unspliced_mapped_segments.sorted error
            [FAILED]
    Error: mapsplice_report failed
    The file mapspliceout/tmp/unsplicedmapped_segments.sorted exists and is not empty.
    When I turn of the --fusion parameter, MapSplice finishes with no errors.

    Has anyone an idea, where the problem might be?
    ecSeq Bioinformatics is Europe’s leading provider of hands-on bioinformatics workshops and professional data analysis in the field of Next-Generation Sequencing (NGS).

  • #2
    I also tried to contact the developer directly (10/25/2012:[email protected]). They haven't answered yet.
    ecSeq Bioinformatics is Europe’s leading provider of hands-on bioinformatics workshops and professional data analysis in the field of Next-Generation Sequencing (NGS).

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