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
- - - - -

How's this RNA


  • Please log in to reply
5 replies to this topic

#1 ram

ram

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 96 posts
0
Neutral

Posted 01 May 2010 - 07:11 AM

This is 1% agarose picture of plant RNA. How is this RNA appearing?...is there any sort of degradation? I have read somewhere that 28s rRNA band should be brighter than 18s rRNA (which is not the case with this pic (?)), otherwise it indicates degradation. Is that true?

Posted Image

Below is the RNA isolated from different stage of tissue...where 28s > 18s

Posted Image

Thanks
Ram

Edited by ram, 01 May 2010 - 07:17 AM.

If you don't know it, then ask it! Better to ask and look foolish to some then not ask and stay stupid.

#2 pDNA

pDNA

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 493 posts
14
Good

Posted 01 May 2010 - 10:34 PM

the RNA appears somehow degraded ...indicated by that smear.

Its hard to tell how much further processing is affected by taking a look at an agarose gel (if you use different exposure the appearance might differ).

We always use the Agilent Bioanalyzer to check the integrity of the RNA ...it gives you a RIN number that measures the quality of the RNA ...the higher the better ...if it is below 7 we do not use the samples for reverse transcription.

How are you going to further process your RNA? ...if am i honest ...my personal feeling towards all this RNA quality stuff is that the efforts made are somehow exaggerated. It always depends on the steps downstream the RNA isolation to what extend the RNA quality is a critical factor.

Regards,
p

#3 ram

ram

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 96 posts
0
Neutral

Posted 01 May 2010 - 10:48 PM

Thanks for the comments p
I am going to use this RNA for reverse transcription followed by real-time PCR
If you don't know it, then ask it! Better to ask and look foolish to some then not ask and stay stupid.

#4 pDNA

pDNA

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 493 posts
14
Good

Posted 01 May 2010 - 11:43 PM

you can go through that paper! maybe it helps you a bit!

I think the most important thing is to take care in the analysis of your results (normalization!!).

Good luck!

Regards,
p

Attached Files



#5 lihkin

lihkin

    member

  • Active Members
  • Pip
  • 10 posts
0
Neutral

Posted 02 May 2010 - 12:14 AM

The RNA quality looks somewhat Ok. I also think that your agarose gel has some problem with solidifying/buffer since it appears a little uneven in run and smudgy. Check the attached picture for a RNA gel 1.2%. we do it all the time in lab so its pretty standard. You can try the expts you have mentioned with this quality and see if it works.
Thanks,
Lihkin

#6 ram

ram

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 96 posts
0
Neutral

Posted 02 May 2010 - 05:42 PM

Thank you for the paper p!
@ lihkin Thanks for the comments; I didn't see any attached picture!
If you don't know it, then ask it! Better to ask and look foolish to some then not ask and stay stupid.




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

©1999-2013 Protocol Online, All rights reserved.