Unconfigured Ad

Collapse
X
 
  • Time
  • Show
Clear All
new posts
  • aaronreba
    Junior Member
    • Jun 2012
    • 2

    #1

    Using Tophat-Fusion to detect Structural Variation in E. coli

    Hello,

    I'm trying to use Tophat-Fusion on a set of E. coli for structural variation detection. The program seems to run with no errors, but the output doesn't seem correct.

    I have a few simple questions first.

    When giving paired end reads to Tophat-Fusion, how should they be passed? Should they be in separate files with similar names like set_of_reads_1.fastq and set_of_reads_2.fastq with corresponding read names for the pairs? Or should they be merged into one file like set_of_reads.fastq such that as the file is being read, every 2 reads is a pair?

    I have built my bowtie files like so:
    bowtie-build REL606.5.gbk bowtie_REL606.5
    Which seems to build correctly.

    My reads are 50 bps in length with a gap size of 100. I then call tophat like so:

    tophat-fusion -p 12 --solexa-quals -r 100 --mate-std-dev 20 -o paired_tophat bowtie_REL606.5 set_1.fastq set_2.fastq

    tophat-fusion -p 12 --solexa-quals -r 100 --mate-std-dev 20 -o merged_tophat bowtie_REL606.5 set.fastq

    I have called it in 2 different ways because I'm unsure of the read method I mentioned above.

    When it completed, I tried examining the sam file but the samtools view command fails with the following error:
    [sam_read1] reference 'REL606.5-REL606.5' is recognized as '*'.
    Parse error at line 2428: invalid CIGAR operation

    How would I be able to examine the sam file?

    Platform: Linux

    Versions:
    TopHat v0.1.0 (Beta)
    bowtie version 0.12.7
    Samtools Version: 0.1.15 (r949:203)

    Thanks
    Last edited by aaronreba; 06-27-2012, 12:48 PM.
  • aaronreba
    Junior Member
    • Jun 2012
    • 2

    #2
    I've found out my questions for anyone else interested. I was apparently using an old version of Tophat. I learned this after downloading only the binaries of Tophat2. The reads also must be in separate files like so:

    File 1:

    read1.1
    atgatgc...
    +
    #$@#$...
    read2.1
    atgatgc...
    +
    #$@#$...

    File 2:

    read1.2
    atgatgc...
    +
    #$@#$...
    read2.2
    atgatgc...
    +
    #$@#$...
    Last edited by aaronreba; 06-28-2012, 10:52 AM.

    Comment

    Latest Articles

    Collapse

    • SEQadmin2
      Beyond CRISPR/Cas9: Understand, Choose, and Use the Right Genome Editing Tool
      by SEQadmin2



      CRISPR/Cas9 sparked the gene editing revolution for both research and therapeutics.1 But this system still showed severe issues that limited its applications. The most prominent were the heavy reliance on PAM sequences, delivery limitations, double-stranded breaks that prompt unintended edits and cell death, and editing inefficiency (both in targeting and in knock-in reliability).

      Despite this, “CRISPR helped turn genome editing from a specialized technique into
      ...
      07-31-2026, 11:01 AM
    • SEQadmin2
      Proteomic Platforms: How to Choose the Right Analytical Strategy to Improve Detection and Clinical Applications
      by SEQadmin2


      Proteomics platforms are evolving rapidly, with advances in mass spectrometry and affinity-based approaches expanding what researchers can detect and at what scale. As the field moves toward deeper proteome coverage and clinical applications, scientists face an increasingly complex landscape of tools. This article will explore how researchers are navigating these choices to find the right platform for their work.

      The systematic characterization of the human proteome has
      ...
      07-20-2026, 11:48 AM
    • SEQadmin2
      Advanced Sequencing Platforms Tackle Neuroscience’s Toughest Genomics Problems
      by SEQadmin2



      Genomics studies in neuroscience face a special challenge due to the brain’s complexity and scarcity of samples. Mapping changes in cell type and state using conventional next-generation sequencing methods remains challenging. Advances in technologies like single-cell sequencing, spatial transcriptomics, and long-read sequencing have opened the door to deeper studies of the brain and diseases like Alzheimer’s, amyotrophic lateral sclerosis (ALS), and schizophrenia.
      ...
      07-09-2026, 11:10 AM

    ad_right_rmr

    Collapse

    News

    Collapse

    Topics Statistics Last Post
    Started by SEQadmin2, 08-03-2026, 10:13 AM
    0 responses
    20 views
    0 reactions
    Last Post SEQadmin2  
    Started by SEQadmin2, 07-31-2026, 02:55 AM
    0 responses
    34 views
    0 reactions
    Last Post SEQadmin2  
    Started by SEQadmin2, 07-24-2026, 12:17 PM
    0 responses
    24 views
    0 reactions
    Last Post SEQadmin2  
    Started by SEQadmin2, 07-23-2026, 11:41 AM
    0 responses
    21 views
    0 reactions
    Last Post SEQadmin2  
    Working...