Unconfigured Ad

Collapse
X
 
  • Filter
  • Time
  • Show
Clear All
new posts
  • mediator
    Member
    • Nov 2010
    • 27

    Tophat paired end alignment help: comma or blank to separate two pairs?

    Hi All,
    I am using the current command to run a PE alignment in Tophat:
    $ tophat -p 4 -m 2 -r 200 /index/hg19 /data/raw/N1_1.fa,N1_2.fa

    my question is, both N1_1.fa and N1_2.fa are the paired end reads generated from a single human tissue sample. Should I use comma or blank to separate those two? And here is the log for the alignment, is 76% alignment looks normal? Thanks!

    # reads processed: 242980185
    # reads with at least one reported alignment: 185782139 (76.46%)
    # reads that failed to align: 56208030 (23.13%)
    # reads with alignments suppressed due to -m: 990016 (0.41%)
    Reported 457696000 alignments to 1 output stream(s)
  • jbrwn
    Member
    • Mar 2011
    • 37

    #2
    a space between file_1 and file_2.

    Comment

    • mediator
      Member
      • Nov 2010
      • 27

      #3
      @jbrwn
      Thanks! Hope that improves my alignment rate.

      Comment

      • jbrwn
        Member
        • Mar 2011
        • 37

        #4
        Originally posted by mediator View Post
        @jbrwn
        Thanks! Hope that improves my alignment rate.
        i typically include --mate-std-dev <int> as well, though i'm not sure how much difference it makes. this information should be available from the bioanalyzer output or someone-from-the-lab output.

        Comment

        Latest Articles

        Collapse

        • GATTACAT
          Reply to Nine Things a Sample Prep Scientist Thinks About Before Sequencing
          by GATTACAT
          Love this - good data definitely starts from good input, and poor input can only give relatively poor data. I particularly like the mention of Nanodrop/absorbance based methods for quantification. It's such a toss up if you'll get an accurate reading or what amounts to a randomly generated number, and a lot of library/sequencing related issues can be traced back to poor quant.
          07-01-2026, 11:43 AM
        • 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

        ad_right_rmr

        Collapse

        News

        Collapse

        Topics Statistics Last Post
        Started by SEQadmin2, 07-02-2026, 11:08 AM
        0 responses
        21 views
        0 reactions
        Last Post SEQadmin2  
        Started by SEQadmin2, 06-30-2026, 05:37 AM
        0 responses
        21 views
        0 reactions
        Last Post SEQadmin2  
        Started by SEQadmin2, 06-26-2026, 11:10 AM
        0 responses
        21 views
        0 reactions
        Last Post SEQadmin2  
        Started by SEQadmin2, 06-17-2026, 06:09 AM
        0 responses
        54 views
        0 reactions
        Last Post SEQadmin2  
        Working...