Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
- - - - -

RNA extraction from flat worms

RNA extraction worms small yield Trizol

  • Please log in to reply
2 replies to this topic

#1 Ksenia



  • Members
  • Pip
  • 2 posts

Posted 27 November 2013 - 01:46 AM

Dear all,

I am extracting RNA from flat worms (Gyrodactylus salaris) with Trizol-based method, but I do not get any RNA, or RNA amounts are very low (the control, fish tissue samples are extracted normally).


 Alive worms were frozen in liquid nitrogen in a drop of water (40 microliters). The number of worms per sample is around 30, and they are quite small (0.1 miliimeters). From this fresh-frozen samples I get nothing, whereas I was able to extract RNA from the same number of RNA-later stored worms previously.


I tried the standard protocol; additionally homogenizing the frozen drop of water to powder with immediate adding of Trizol; letting the samples to sit longer in Trizol for better lyzation; elongating the RNA precipitation step, but nothing seems to work. 


Does anybody have an experience with extracting from very small initial amounts of tissue or may suggest something?


​I will appreciate your help!!


#2 phage434



  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 2,747 posts

Posted 27 November 2013 - 06:50 AM

You could try flash freezing and pulverizing. Best would probably be freeziing in LN2 or dry ice/ethanol,  followed by grinding with a bead beater in a small volume of trizoi. I would add the trizol directly to the frozen sample pellet before the grinding.

#3 Ksenia



  • Members
  • Pip
  • 2 posts

Posted 28 November 2013 - 05:30 AM

Thank you! Actually, that what I did initially, with no success however..

Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.