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  • sweetph3
    Member
    • Jun 2013
    • 22

    #1

    Strange RNA Bioanalyzer trace, any thoughts?

    We have only just started using the Bioanalyzer RNA 6000 Nano chips, and I haven't seen a good trace yet. Today's was very strange....

    There is a main peak at ~40 seconds that blows the rest of the peaks away (one was over 1000 FU). The software is calling it 18S, but there isn't a corresponding 28S peak.

    I've attached a PDF of the four traces.

    I loaded 1 ul of total RNA, with concentrations of 7.2 to 31.6 ng/ul as measured by Qubit. These samples have been in -80 since May (2 months), made on 5/8/14 and thawed once on 5/9/14 to make TruSeq RNA libraries (which worked beautifully). Any thoughts on that huge peak? Thank you for your insight!
    Attached Files
  • MU Core
    Member
    • Apr 2008
    • 60

    #2
    The profile is reminiscent of insect RNA that has the characteristic of the 28S breaking in to two subunits when heated. This is a phenomenon observed for many years. The attached articles demonstrates discusses this in honeybee RNA.
    Attached Files

    Comment

    • nucacidhunter
      Jafar Jabbari
      • Jan 2013
      • 1250

      #3
      MU Core comments are correct. This topic has also been discussed here: http://seqanswers.com/forums/showthread.php?t=3898

      Comment

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