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

miRNA isolation from liquid samples


  • Please log in to reply
18 replies to this topic

#16 Fizban

Fizban

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 63 posts
1
Neutral

Posted 16 June 2010 - 01:40 AM

Hi again,

Forgot to ask, how much TRIZOL do you guys use to extract from the serum ???


I use 1 mL for each 400 ul of serum. then use 1ml of isopropanol after adding glycogen then proceed as indicated by standard protocol
bye
Fiz

#17 Bastard

Bastard

    member

  • Members
  • Pip
  • 1 posts
0
Neutral

Posted 24 June 2010 - 06:03 AM

Forget about nanoDrop or picoDrop its crap for this!

Try to quantify your RNA with Bioanalyser or the Experion RNA (highsense) Chip (even though desalting is needed)! Maybe the concentration is loads lower than you expect!

#18 Fizban

Fizban

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 63 posts
1
Neutral

Posted 24 June 2010 - 09:37 PM

Forget about nanoDrop or picoDrop its crap for this!

Try to quantify your RNA with Bioanalyser or the Experion RNA (highsense) Chip (even though desalting is needed)! Maybe the concentration is loads lower than you expect!


Forget Experion too! the only one is bioanalyzer but still it's hard. what i'd like to know is how many people published papers in which they quantitate RNA with nanodrop. beats me ;)
Fiz

#19 SAN1017

SAN1017

    member

  • Members
  • Pip
  • 4 posts
0
Neutral

Posted 23 July 2010 - 01:11 AM

I also trying to extract the miRNA in plasma.
I use the trizol + Qiagen's RNeasy plus kit.
For about 500ul plasma, in 30ul elution, the Nanodrop reading is about 10-15 ng/ul. The 260/280 is alright, but also 230/260 is no gd.
However, it is no problem for the real-time PCR result.

But I would like to ask, this reading is the total RNA but not 100% miRNA, right?
Furthermore, as i know, the circulating miRNA is inside the exosome. Trizol enough to break down the exosome and release the miRNA for purification?

San




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

©1999-2013 Protocol Online, All rights reserved.