Unconfigured Ad

Collapse
X
 
  • Filter
  • Time
  • Show
Clear All
new posts
  • ibn.adam
    Junior Member
    • Aug 2011
    • 8

    coverageBed: command not found

    Dear all,

    I am mapping one end mRNA sequencing on TOPHAT with human reference genome hg19.

    It looks like script worked partially and It has generated .BAM file of 7 million MB. But geneexpression.txt are empty.

    Here is the error file:


    [Fri Oct 28 18:13:55 2011] Beginning TopHat run (v1.3.0)
    -----------------------------------------------
    [Fri Oct 28 18:13:55 2011] Preparing output location /home/ah644/tophat_output/intestinal/mRNA/C1055/
    [Fri Oct 28 18:13:55 2011] Checking for Bowtie index files
    [Fri Oct 28 18:13:55 2011] Checking for reference FASTA file
    [Fri Oct 28 18:13:55 2011] Checking for Bowtie
    Bowtie version: 0.12.7.0
    [Fri Oct 28 18:13:55 2011] Checking for Samtools
    Samtools Version: 0.1.17
    [Fri Oct 28 18:13:55 2011] Generating SAM header for /home/ah644/hg19andhs37/hg19
    [Fri Oct 28 18:13:56 2011] Preparing reads
    format: fastq
    quality scale: phred33 (default)
    [Fri Oct 28 18:13:56 2011] Reading known junctions from GTF file
    Left reads: min. length=76, count=151037396
    [Fri Oct 28 19:33:26 2011] Mapping left_kept_reads against hg19 with Bowtie
    [Fri Oct 28 21:46:41 2011] Processing bowtie hits
    [Fri Oct 28 23:43:18 2011] Mapping left_kept_reads_seg1 against hg19 with Bowtie (1/3)
    [Sat Oct 29 00:38:57 2011] Mapping left_kept_reads_seg2 against hg19 with Bowtie (2/3)
    [Sat Oct 29 01:34:48 2011] Mapping left_kept_reads_seg3 against hg19 with Bowtie (3/3)
    [Sat Oct 29 02:21:31 2011] Searching for junctions via segment mapping
    [Sat Oct 29 05:17:18 2011] Retrieving sequences for splices
    [Sat Oct 29 05:22:36 2011] Indexing splices
    [Sat Oct 29 05:23:14 2011] Mapping left_kept_reads_seg1 against segment_juncs with Bowtie (1/3)
    [Sat Oct 29 05:48:31 2011] Mapping left_kept_reads_seg2 against segment_juncs with Bowtie (2/3)
    [Sat Oct 29 06:11:28 2011] Mapping left_kept_reads_seg3 against segment_juncs with Bowtie (3/3)
    [Sat Oct 29 06:28:37 2011] Joining segment hits
    [Sat Oct 29 08:53:02 2011] Reporting output tracks
    -----------------------------------------------
    Run complete [16:42:32 elapsed]
    /opt/torque/mom_priv/jobs/80964.bulldogn-rocks.wss.yale.edu.SC: line 15: coverageBed: command not found


    Pipeline:

    #!/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/ah644/projects
    cd $ROOT/$SAMPLE
    ROOTB=/home/ah644/tophat_output/
    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 -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

Latest Articles

Collapse

  • SEQadmin2
    Nine Things a Sample Prep Scientist Thinks About Before Sequencing
    by SEQadmin2


    I’m not a sequencing expert. I’m a purification scientist who uses NGS to evaluate workflows my group develops. With this perspective, we think about the sample first and the NGS workflow second. The sequencer is an exceptionally honest reporter, but it can only report on what you give it, so whether you get clean, interpretable data from an NGS workflow is largely determined before you begin.


    Here are nine questions we think about, in roughly the order they matter, before...
    06-18-2026, 07:11 AM
  • SEQadmin2
    From Collection to Sequencing: Why Sample Preparation and Preservation Define Sequencing Data
    by SEQadmin2


    Data variability is still an issue in sequencing technologies despite the advances in reproducibility and accuracy of these platforms. But the problem does not originate in the sequencing itself, but in the previous steps, before the sample reaches the sequencer.


    The first step is collection, followed by preservation and sample preparation for analysis. Most scientists overlook those steps, but not being careful might just be skewing the experiment’s results.
    ...
    06-02-2026, 10:05 AM
  • SEQadmin2
    Single-Cell Sequencing at an Inflection Point: Early Impacts of New Platforms and Emerging Trends
    by SEQadmin2


    With the launch of new single-cell sequencing platforms in 2026, the field stands at an exciting inflection point. This article surveys the most impactful advances in the field and discusses how they’re reshaping research in cancer, immunology, and beyond.


    Introduction

    Single-cell sequencing technologies have undergone remarkable advances over the past decade, transitioning from low-throughput experimental approaches to highly scalable platforms capable of...
    05-22-2026, 06:42 AM

ad_right_rmr

Collapse

News

Collapse

Topics Statistics Last Post
Started by SEQadmin2, 06-17-2026, 06:09 AM
0 responses
24 views
0 reactions
Last Post SEQadmin2  
Started by SEQadmin2, 06-09-2026, 11:58 AM
0 responses
40 views
0 reactions
Last Post SEQadmin2  
Started by SEQadmin2, 06-05-2026, 10:09 AM
0 responses
47 views
0 reactions
Last Post SEQadmin2  
Started by SEQadmin2, 06-04-2026, 08:59 AM
0 responses
49 views
0 reactions
Last Post SEQadmin2  
Working...