Seqanswers Leaderboard Ad

Collapse

Announcement

Collapse
No announcement yet.
X
 
  • Filter
  • Time
  • Show
Clear All
new posts

  • TopHat Error: you must set the mean inner distance between mates with -r?

    I tried to use TOPHAT on Arabidopsis mRNA-seqs, they are not pair end reads. I've run the example data you provided, so the installation is all right.
    For my own data, I've built bowtie index "tair9_bwt" for Arabidopsis genome sequences. I have 6 fastq files of mRNA-seqs. I want to use TOPHAT to find new transcript boundaries in intergenic regions.

    I used the following command to test two of the six fq files, it could generate the 3 output files successfully.
    tophat ~/bowtie-0.12.5/tair9_bwt/tair9_bwt tail.fastq,test1.fastq

    But when I tried to use options like -i -I or -G or other options, it failed:
    -----------------------
    $ tophat ~/bowtie-0.12.5/tair9_bwt/tair9_bwt tail.fastq,test1.fastq -i 30 -I 15000
    Error: you must set the mean inner distance between mates with -r
    Traceback (most recent call last):
    File "/home/wux3/mrnaseq/tophat/bin/tophat", line 1635, in ?
    sys.exit(main())
    File "/home/wux3/mrnaseq/tophat/bin/tophat", line 1535, in main
    exit(1)
    TypeError: 'str' object is not callable
    ----------------------

    The description of -r option is:
    -r/--mate-inner-dist <int> This is the expected (mean) inner distance between mate pairs. For, example, for paired end runs with fragments selected at 300bp, where each end is 50bp, you should set -r to be 200. There is no default, and this parameter is required for paired end runs.

    But the mRNA-seqs I have are not paired end, anybody knows how to set this option on my data? And, why the "-r" option is not neccessary when no options used in the commandline?

    Thank you in advance.

  • #2
    Now, it is OK. Just used options before the fastq... -.-

    tophat -G TAIR9_GFF3_tophat -i 30 -I 15000 ~/bowtie-0.12.5/tair9_bwt/tair9_bwt tail.fastq,head.fastq
    Last edited by xhuister; 04-28-2010, 01:08 PM.

    Comment


    • #3
      thanks for posting this, just saved me a lot of frustrations...

      Comment

      Latest Articles

      Collapse

      • seqadmin
        Current Approaches to Protein Sequencing
        by seqadmin


        Proteins are often described as the workhorses of the cell, and identifying their sequences is key to understanding their role in biological processes and disease. Currently, the most common technique used to determine protein sequences is mass spectrometry. While still a valuable tool, mass spectrometry faces several limitations and requires a highly experienced scientist familiar with the equipment to operate it. Additionally, other proteomic methods, like affinity assays, are constrained...
        04-04-2024, 04:25 PM
      • seqadmin
        Strategies for Sequencing Challenging Samples
        by seqadmin


        Despite advancements in sequencing platforms and related sample preparation technologies, certain sample types continue to present significant challenges that can compromise sequencing results. Pedro Echave, Senior Manager of the Global Business Segment at Revvity, explained that the success of a sequencing experiment ultimately depends on the amount and integrity of the nucleic acid template (RNA or DNA) obtained from a sample. “The better the quality of the nucleic acid isolated...
        03-22-2024, 06:39 AM

      ad_right_rmr

      Collapse

      News

      Collapse

      Topics Statistics Last Post
      Started by seqadmin, 04-11-2024, 12:08 PM
      0 responses
      24 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 04-10-2024, 10:19 PM
      0 responses
      25 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 04-10-2024, 09:21 AM
      0 responses
      21 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 04-04-2024, 09:00 AM
      0 responses
      52 views
      0 likes
      Last Post seqadmin  
      Working...
      X