Unconfigured Ad

Collapse
X
 
  • Time
  • Show
Clear All
new posts
  • Seqanalysis
    Junior Member
    • May 2013
    • 2

    #1

    Completing genome of thermophilic bacteria

    Hi,
    I sequenced the genome of a thermophilic bacteria using Illumina. There was a gap of 37kb that was missed during sequencing but I was able to fill that using sanger sequecing. This region seemed to be highly conserved but has 33genes that are designated as hypothetical proteins.
    My problem is I am not able to sequence a gene which is about 3.8kb completely. With internal pair of primers i was able to get 1.9kb sequence but am not able to walk in using primers from neighboring genes which is strange as I know the genes are together.
    Can anyone suggest me what could be the possibilities?

    This region is probably not stable in this organism it seems to be susceptible to degradation during extraction.
    What could be the reasons?
  • ECO
    --Site Admin--
    • Oct 2007
    • 1360

    #2
    Super high GC? Repetitive elements? Gene family? N4-mC content interfering with PCR?

    Sounds like a job for a couple PacBio SMRTcells*


    *disclaimer: I used to work there. But this is textbook RS/RSII.

    Comment

    • Seqanalysis
      Junior Member
      • May 2013
      • 2

      #3
      GC is 46% for my organism. Surprisingly I get the pcr products in other species that I use as control. This region is highly conserved in this genus. Repetitive seq yes they have a lot but then again it's the same in other species.

      I really appreciate your suggestions. I would be glad to know more

      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, 10-15-2026, 10:52 AM
      0 responses
      21 views
      0 reactions
      Last Post SEQadmin2  
      Started by SEQadmin2, 10-14-2026, 05:39 AM
      0 responses
      26 views
      0 reactions
      Last Post SEQadmin2  
      Started by SEQadmin2, 10-13-2026, 09:56 AM
      0 responses
      22 views
      0 reactions
      Last Post SEQadmin2  
      Started by SEQadmin2, 10-09-2026, 11:14 AM
      0 responses
      28 views
      0 reactions
      Last Post SEQadmin2  
      Working...