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 clean up - what is the rna recovery


  • Please log in to reply
2 replies to this topic

#1 wntiong

wntiong

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 58 posts
0
Neutral

Posted 28 September 2009 - 08:59 PM

Hi guys, i am isolating rna from whole blood using trizol method, and some of my samples have quite good A260/280 and A260/230 ratio of btw 1.7 to 2.0. In fact, i think it is necessary for me to do a rna clean up using spin column method (Qiaamp rna blood kit), but i am not sure what is the rna recovery % since i only managed to get about 10 ug of rna after trizol extraction, i afraid it will get much lesser after clean up step. FYI, i need at least 10ug for microarray work. Kindly advise. Thanks

#2 Dr Teeth

Dr Teeth

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 221 posts
1
Neutral

Posted 29 September 2009 - 03:31 AM

Hi guys, i am isolating rna from whole blood using trizol method, and some of my samples have quite good A260/280 and A260/230 ratio of btw 1.7 to 2.0. In fact, i think it is necessary for me to do a rna clean up using spin column method (Qiaamp rna blood kit), but i am not sure what is the rna recovery % since i only managed to get about 10 ug of rna after trizol extraction, i afraid it will get much lesser after clean up step. FYI, i need at least 10ug for microarray work. Kindly advise. Thanks


Microarray work is much more sensitive to contaminants in your RNA sample than qRT-PCR. You should do a spin column cleanup. If your recovery is too low, consider pooling like samples.

Science is simply common sense at its best that is rigidly accurate in observation and merciless to fallacy in logic.
Thomas Henry Huxley

#3 wntiong

wntiong

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 58 posts
0
Neutral

Posted 29 September 2009 - 11:02 PM

Hi guys, i am isolating rna from whole blood using trizol method, and some of my samples have quite good A260/280 and A260/230 ratio of btw 1.7 to 2.0. In fact, i think it is necessary for me to do a rna clean up using spin column method (Qiaamp rna blood kit), but i am not sure what is the rna recovery % since i only managed to get about 10 ug of rna after trizol extraction, i afraid it will get much lesser after clean up step. FYI, i need at least 10ug for microarray work. Kindly advise. Thanks


Microarray work is much more sensitive to contaminants in your RNA sample than qRT-PCR. You should do a spin column cleanup. If your recovery is too low, consider pooling like samples.


I did the spin column, the band is much clearer now, is just that the amount of rna is half of the amount i obtained after trizol. How to pool my samples during spin column cleanup?




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

©1999-2013 Protocol Online, All rights reserved.