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  • Tophat colorspace problem --integer-quals 0

    Hello everyone,

    Is there an tophat option for setting the --integer-quals to 0 when using colorspace reads? I have a fastq file with phred33 quality. According to the manual when using the --color (or -C) option, the default of --integer-qual is set to 'yes'. There is no option to set the --integer-quals to zero, when I try --integer-qual 0, I retrieve the error: Error: Could not find Bowtie index files (0.*.ebwt).

    When executing tophat, this is my output, which many people on this forum have when using colorspace, but noone has a solution:
    Code:
    [2013-03-18 10:42:01] Beginning TopHat run (v2.0.6)
    -----------------------------------------------
    [2013-03-18 10:42:01] Checking for Bowtie
                      Bowtie version:        0.12.8.0
    [2013-03-18 10:42:01] Checking for Samtools
                    Samtools version:        0.1.14.0
    [2013-03-18 10:42:01] Checking for Bowtie index files
    [2013-03-18 10:42:01] Checking for Bowtie index files
    [2013-03-18 10:42:01] Checking for reference FASTA file
    [2013-03-18 10:42:01] Generating SAM header for /home/sge_share_fedor12/jetse/GENOMES/GRCh37_gatk/GRCh37_gatk_colorspace
            format:          fastq
            quality scale:   phred33 (default)
    [2013-03-18 10:42:16] Reading known junctions from GTF file
    [2013-03-18 10:42:42] Preparing reads
             left reads: min. length=49, max. length=49, 99681 kept reads (319 discarded)
    [2013-03-18 10:42:44] Using pre-built transcriptome index..
    [2013-03-18 10:42:56] Mapping left_kept_reads to transcriptome human_GRCh37_gatk_index with Bowtie 
            [FAILED]
    Error running bowtie:
    Too few quality values for read: 7T0@
            are you sure this is a FASTQ-int file?
    Command: /usr/bin/bowtie -q -C --col-keepends -v 3 -k 1 -m 1 -S -p 1 --sam-nohead --max /dev/null /home/sge_share_fedor12/jetse/GENOMES/GRCh37_gatk/human_GRCh37_gatk_index -
    When reading this output, Tophat recognized the phred33 quality scale but it wants a FASTQ-int file...
    Anyone knows how to solve this error?

    [EDIT]
    Tried to use tophat with the --integer-quals, but then already stucking at prep_reads: terminate called after throwing an instance of 'int'

    [EDIT]
    Now tried to convert the fastq reads to integer-quals quality's, but then I get the error:
    Code:
    Error encountered parsing file /home/sge_share_fedor12/jetse/wntSignalling/test/splitted/p1.A3754971/p1.A3754971_0_F3.fastq:
     Length mismatch between sequence and quality strings for A3754971_F3_0001_0 (49 vs 146).
    Did anyone use fastq files from a solid sequencer on tophat? How did your files look?
    Last edited by Jetse; 03-18-2013, 06:50 AM. Reason: Tried some more options

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