Unconfigured Ad

Collapse
X
 
  • Time
  • Show
Clear All
new posts
  • nasepu
    Junior Member
    • Sep 2012
    • 5

    #1

    Interrogate .fq file for specific sequence?

    I found a very suspect major snp when digging through my sequence data, identical to a construct I've previously made in the lab. I'm concerned this is contamination, and it has too significant a phenotype to be ignored. The best way I could think to check if the plasmid somehow made its way in is to check for other stuff that would be on a plasmid, like ampicillin or kanamycin resistance.

    I'm unsure what the best way to go about this is. I briefly tried aligning the entire .fq file against the amp gene sequence, which threw an error that can probably be fixed, but the whole idea of trying to align billions of reads against one tiny gene rubbed me the wrong way. Is there a better way to go about this/any tools that would be more useful?
  • Bukowski
    Senior Member
    • Jan 2010
    • 388

    #2
    On the grounds of 'right tool for the right job' using an aligner specifically designed to align ngs reads to a sequence seems like the right way to go to me, as it's likely to be faster than (e.g.) BLAT.

    With such a small search space it doesn't seem like the job would take that long - why not try aligning to the vector sequence not just the gene?

    Comment

    • swbarnes2
      Senior Member
      • May 2008
      • 910

      #3
      Try:

      Code:
      grep AGTTAGCTCTCGATAGGCTAT read1.fq
      For whatever sequence you are looking for.

      Comment

      Latest Articles

      Collapse

      • SEQadmin2
        Multiomic Liquid Biopsy: What DNA/RNA Panels Reveal That DNA Alone Can’t
        by SEQadmin2



        Tumors contain complex mixtures of cell populations that evolve in response to treatment and host-specific factors. Tissue biopsies provide only static snapshots of a tumor’s state. But what if we could follow that evolution over time using a minimally invasive technique?

        Liquid biopsies can detect tumor-derived material circulating in the bloodstream, providing a way to study treatment response, detect emerging resistance, and discover new biomarkers. Circulating
        ...
        10-14-2026, 09:55 AM
      • SEQadmin2
        New Genomics Technologies Take Aim at Long-Standing Limits
        by SEQadmin2


        Researchers using sequencing and genomics tools often have to make trade-offs. They can choose between speed or scale, short reads or long-range information, or targeted panels or a view of the whole transcriptome. New technologies that have been released this year are built to address those tough choices.

        We asked six companies the same four questions to learn about their latest products. The new technologies bring a lot to the table, including rethinking sequencing
        ...
        09-28-2026, 10:25 AM

      ad_right_rmr

      Collapse

      News

      Collapse

      Topics Statistics Last Post
      Started by SEQadmin2, Yesterday, 11:13 AM
      0 responses
      19 views
      0 reactions
      Last Post SEQadmin2  
      Started by SEQadmin2, Yesterday, 10:48 AM
      0 responses
      18 views
      0 reactions
      Last Post SEQadmin2  
      Started by SEQadmin2, 10-15-2026, 10:52 AM
      0 responses
      36 views
      0 reactions
      Last Post SEQadmin2  
      Started by SEQadmin2, 10-14-2026, 05:39 AM
      0 responses
      42 views
      0 reactions
      Last Post SEQadmin2  
      Working...