Seqanswers Leaderboard Ad

Collapse

Announcement

Collapse
No announcement yet.
X
 
  • Filter
  • Time
  • Show
Clear All
new posts

  • No 28S peak

    Hey,

    I am using Direct-zol™ RNA MiniPrep to isolate my total RNA from cells in TRIzol. I did isolate it and run bioanalyzer - results in attachment. I am clearly missing 28S peak, material is coming from M.lignano, flatworm. Any ideas? Bioanalyzer also gives an error: unexpected signal at 5S. Is my RNA degraded? Thanks a lot!
    Attached Files

  • #2
    While I know nothing about your organism, I do know that if you denature drosophila RNA the peaks are virtually on top of each other-more like a doublet than two distinct peaks-whereas if you don't denature it they resolve better.

    Maybe try running it non-denatured and see what you get?

    Comment


    • #3
      "material is coming from M.lignano" - Do you mean Macrostomum lignano, a flatworm? I recently extracted RNA from another flatworm, Opistorchis, and it looks exactly the same.
      THis is actually not rare for invertebrates, i've seen the same for snails, amphipods etc.
      Attached Files

      Comment


      • #4
        Originally posted by MLog View Post
        "material is coming from M.lignano" - Do you mean Macrostomum lignano, a flatworm? I recently extracted RNA from another flatworm, Opistorchis, and it looks exactly the same.
        THis is actually not rare for invertebrates, i've seen the same for snails, amphipods etc.
        Yes, I am working with flatworms. I indeed see that more often while testing other flatworm species. In denaturing conditions the bigger subunit dissociates and forms two smaller, which seems to overlap with 18S. Thanks for the feedback!

        Comment


        • #5
          Originally posted by kwaraska View Post
          While I know nothing about your organism, I do know that if you denature drosophila RNA the peaks are virtually on top of each other-more like a doublet than two distinct peaks-whereas if you don't denature it they resolve better.

          Maybe try running it non-denatured and see what you get?
          Thanks a lot! You are totally correct. We confirmed that with running both: my total RNA and Drospohila RNA on the gel. Peaks looks similar too. Thanks again!

          Comment

          Latest Articles

          Collapse

          • seqadmin
            Exploring the Dynamics of the Tumor Microenvironment
            by seqadmin




            The complexity of cancer is clearly demonstrated in the diverse ecosystem of the tumor microenvironment (TME). The TME is made up of numerous cell types and its development begins with the changes that happen during oncogenesis. “Genomic mutations, copy number changes, epigenetic alterations, and alternative gene expression occur to varying degrees within the affected tumor cells,” explained Andrea O’Hara, Ph.D., Strategic Technical Specialist at Azenta. “As...
            07-08-2024, 03:19 PM
          • seqadmin
            Exploring Human Diversity Through Large-Scale Omics
            by seqadmin


            In 2003, researchers from the Human Genome Project (HGP) announced the most comprehensive genome to date1. Although the genome wasn’t fully completed until nearly 20 years later2, numerous large-scale projects, such as the International HapMap Project and 1000 Genomes Project, continued the HGP's work, capturing extensive variation and genomic diversity within humans. Recently, newer initiatives have significantly increased in scale and expanded beyond genomics, offering a more detailed...
            06-25-2024, 06:43 AM

          ad_right_rmr

          Collapse

          News

          Collapse

          Topics Statistics Last Post
          Started by seqadmin, 07-10-2024, 07:30 AM
          0 responses
          29 views
          0 likes
          Last Post seqadmin  
          Started by seqadmin, 07-03-2024, 09:45 AM
          0 responses
          201 views
          0 likes
          Last Post seqadmin  
          Started by seqadmin, 07-03-2024, 08:54 AM
          0 responses
          212 views
          0 likes
          Last Post seqadmin  
          Started by seqadmin, 07-02-2024, 03:00 PM
          0 responses
          193 views
          0 likes
          Last Post seqadmin  
          Working...
          X