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!
Photo
- - - - -

RNA extraction for RNAseq

rna rnaseq rna extraction

  • Please log in to reply
3 replies to this topic

#1 Tara Burns

Tara Burns

    member

  • Members
  • Pip
  • 3 posts
0
Neutral

Posted 21 August 2015 - 08:14 AM

Hello all,

 

I am trying to extract RNA from embryonic mouse hearts.  I have a few questions, what are people's methods of disruption/homogenization-we don't have any tools for that right now, I am wondering if there is anything cheap I can buy.  Also, while I do the dissections, is it possible to put the tissue immediately in the RLT that comes with the Qiagen kit?  I tried putting it in RNA later, but when I tried to homogenize with a tube mortar and pestle, I couldn't grind the tissue because it got so hard in the RNA later solution.  And finally, do people find that the trizol method is better than the RNA kits from qiagen, and if so does anyone have a protocol?

 

Tara



#2 mdfenko

mdfenko

    an elder emeritus

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 3,294 posts
204
Excellent

Posted 24 August 2015 - 08:40 AM

you can freeze the heart in liquid nitrogen (before or after treating with rnalater) and then pulverize it with either a mortar and pestle or a pulverizer specifically designed for this.


talent does what it can
genius does what it must
i do what i get paid to do

#3 toptea

toptea

    member

  • Active Members
  • Pip
  • 12 posts
0
Neutral

Posted 07 September 2015 - 05:12 PM

I had used IKA polytron and mortar and pestle for disruption and homogenization of a range of tissues for RNA extraction, both results were equally good.

you can put the tissue directly into buffer RLT making sure you proceed further for homogenizing, once you homogenize it , you can store it at -80 for further steps of RNA extraction although its better to finish RNA extraction at the same time. To my experience, kits are easy and sure way for the beginners to get RNA from tissues and Qiagen kit surely works.



#4 BMF

BMF

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 167 posts
12
Good

Posted 08 September 2015 - 10:40 AM

My favorite way is this:

 

Liquid Nitrogen Cooled  Mortar & Pestle

http://www.braintree...?number=3726001

 

Some image for how to do it:

 

https://www.google.c...wIVBF2SCh0VjwJu

 

Then mix the powder of tissue with Trizol or any solution from any KIT for RNA extraction.


Edited by BMF, 08 September 2015 - 10:42 AM.






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

©1999-2013 Protocol Online, All rights reserved.