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  • ibn.adam
    Junior Member
    • Aug 2011
    • 8

    #1

    single end RNA sequence mapping-TOPHAT- noBAM file?

    Dear all,

    I am maping one end mRNA sequencing on TOPHAT with human reference genome hg19. From sequencinglab i had got many fastq files that, I have merged to a sequence.txt file. I am using that sequence.txt file to run on TOPHAT for mapping.

    The problem is, I am not getting BAM file in TOPHAT output. All i am getting is a hg19.fas file, which i believe is the reference genome file I am using.



    I am using following script for TOPHAT.

    #!/bin/bash
    # usage: qsub -v SAMPLE="name" intestinaltophat1lane.pbs
    #PBS -l walltime=100:00:00
    #PBS -m ae
    #PBS -M [email protected]
    ### Set the queue name, given to you when you get a reservation.
    #PBS -l nodes=1: ppn=8
    ROOT=/home/xxx/projects/intestinal/mRNA/
    cd $ROOT/$SAMPLE
    ROOTB=/home/xxx/tophat_output/intestinal/mRNA/
    TOPHATOUTPUT=$ROOTB$SAMPLE
    mkdir $TOPHATOUTPUT
    SEQFILES=`ls *_sequence.txt`
    tophat -p 8 -r 50 --microexon-search -G /home/ah644/hg19andhs37/Homo_sapiens.GRCh37.62.gtf --solexa1.3-quals -o $TOPHATOUTPUT /home/ah644/hg19andhs37/hg19 $SEQFILES
    coverageBed -abam $TOPHATOUTPUT/accepted_hits.bam -b /home/ah644/hg19andhs37/hg19ensembl.BED > $TOPHATOUTPUT/geneexpression.txt
    cut -f 1,2,3,4,13,14,15,16 $TOPHATOUTPUT/geneexpression.txt > $TOPHATOUTPUT/temp
    cut -f 5 /home/ah644/hg19andhs37/hg19ensemblgeneinfo.txt > $TOPHATOUTPUT/temp2
    paste $TOPHATOUTPUT/temp $TOPHATOUTPUT/temp2 > $TOPHATOUTPUT/reducedgeneexpression.txt
    rm $TOPHATOUTPUT/temp*





    please help me in correcting the script.
    many thanks,
    Adam
    Last edited by ibn.adam; 10-28-2011, 06:51 AM.

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