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

Experience with mRNA isolation kits?

mRNA isolation kit RT-PCR

  • Please log in to reply
5 replies to this topic

#1 sarabell

sarabell

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 04 January 2012 - 06:04 AM

Hello,

Does anyone have suggestions for mRNA isolation kits? What kits have you used and how was the quality?

Any advice is appreciated.

#2 BioMiha

BioMiha

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 225 posts
8
Neutral

Posted 04 January 2012 - 11:04 PM

For total RNA isolation we have used RNeasy and it works great. Also used Promega total RNA extraction kit with far less yield. Not sure about kits for mRNA only but I hope this helps.

#3 mdfenko

mdfenko

    an elder

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 2,615 posts
118
Excellent

Posted 05 January 2012 - 11:49 AM

i think this is the one i used (a long time ago, i was building a library) and it worked well:

clontech

you can also check out this page from life technologies:

mrna extraction

Edited by mdfenko, 05 January 2012 - 11:50 AM.

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

#4 ReResearcher

ReResearcher

    member

  • Active Members
  • Pip
  • 8 posts
0
Neutral

Posted 10 January 2012 - 08:17 PM

Typically trizol works well. 70-100mg of tissue will yield a LOT of mRNA.

RNaqueous kits from Ambion work beautifully if you have difficult tissues (especially high fat content ones). I have gotten great results from 2mg pieces of brain and from adipose, which doesnt have much RNA. There is a micro kit (1-10mg) and the regular kit with columns that go to 100mg of tissue. Homogenize then spin down any debris that might clog the filter. Generally the RNA is very pure but (nanodrop readings at 2 for both ratios) but sometimes there will be contamination/carryover, which is fixed with an overnight lithium chloride precipitation. For microarray (or anything you need super clean RNA for) do a DNAase treatment then LiCl precip (both come with the micro kit, LiCl comes with the regular kit).

#5 fiat29

fiat29

    member

  • Members
  • Pip
  • 3 posts
0
Neutral

Posted 23 November 2012 - 01:33 AM

I used combination TRIZOL with RNeasy, it works well, I get high concentration and purity. Just take the aqueus phase after tizol treatment, and then transfer it to new tube, wash by 1 x volume of 70% ethanol, and transfer to the collumn. And you can go on with the normal procedure of RNeasy. Just skip homogenize the tissue with RLT buffer.

#6 memari

memari

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 148 posts
11
Good

Posted 24 November 2012 - 05:35 PM

I have used RNAzol too. It is cheaper that Trizol. (100 ml is 110$ if you buy it from following site or 150$ if you buy it from its distributors, but TRIzol is 233$).

http://www.mrcgene.com/rnazol.htm
-----
Babak Memari





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

©1999-2013 Protocol Online, All rights reserved.