Unconfigured Ad

Collapse
X
 
  • Filter
  • Time
  • Show
Clear All
new posts
  • monkey_SEQ
    Junior Member
    • Oct 2011
    • 6

    High molecular bands in RNA isolation sample

    I am attempting a de novo transcriptome assembly and have a question about genomic DNA contamination. I have treated my RNA samples twice with DNAseI (on-column and then afterwards in solution as it seemed that there was still HMW smears present). After both these treatments I still feel that I have not gotten rid of the HMW smears (see att 1% agarose gel, RNA ladder). I don’t have primers available to definitively test for gDNA. Could somebody please comment on what these HMW smear can be?

    (PS. Only a single rRNA band present as this is an insect species with a ‘hidden break' in the 28S rRNA).

    What effect will contaminated gDNA sequences have on the transcriptome assembly process?

    Much appreciated
    Attached Files
  • ywg
    Junior Member
    • Feb 2012
    • 1

    #2
    How about do a RNAase test. treat small aliquot of rRNA with RNAase. compare with and without RNAase to confirm it is real gDNA contamination?

    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, Today, 11:05 AM
    0 responses
    7 views
    0 reactions
    Last Post SEQadmin2  
    Started by SEQadmin2, 07-02-2026, 11:08 AM
    0 responses
    28 views
    0 reactions
    Last Post SEQadmin2  
    Started by SEQadmin2, 06-30-2026, 05:37 AM
    0 responses
    27 views
    0 reactions
    Last Post SEQadmin2  
    Started by SEQadmin2, 06-26-2026, 11:10 AM
    0 responses
    26 views
    0 reactions
    Last Post SEQadmin2  
    Working...