Unconfigured Ad

Collapse
X
 
  • Filter
  • Time
  • Show
Clear All
new posts
  • PJohn
    Junior Member
    • Jul 2015
    • 2

    NextSeq reads count

    Hi all,

    I counted the number of reads in the same sample in 2 different ways:

    a) On the sequencer (NextSeq), by counting the number of reads identified that passed filter
    b) On the server, in the fastq files using the command wc –l (total lines) and then dividing the result by 4

    The problem is that those two numbers do not match. Shall they match or not? What is the difference? My objective is to get around 50 million reads per sample and I am figuring out which counting option use. Thanks!

    PJ
  • GenoMax
    Senior Member
    • Feb 2008
    • 7142

    #2
    Are you doing adapter trimming on the sequencer? I assume the difference may be explained by reads that passed filter but were trimmed due to presence of adapter/dimers.

    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
    22 views
    0 reactions
    Last Post SEQadmin2  
    Started by SEQadmin2, 06-30-2026, 05:37 AM
    0 responses
    23 views
    0 reactions
    Last Post SEQadmin2  
    Started by SEQadmin2, 06-26-2026, 11:10 AM
    0 responses
    22 views
    0 reactions
    Last Post SEQadmin2  
    Started by SEQadmin2, 06-17-2026, 06:09 AM
    0 responses
    55 views
    0 reactions
    Last Post SEQadmin2  
    Working...