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 isolation


  • Please log in to reply
5 replies to this topic

#1 anonymous

anonymous

    Veteran

  • PipPipPipPipPipPipPipPipPipPip
  • 1,890 posts
1
Neutral

Posted 25 January 2001 - 10:00 PM

while extracting RNA, we always get low 260/280 ratio about 1.4-1.5 . is there any better way than repeating our protocol?

#2 anonymous

anonymous

    Veteran

  • PipPipPipPipPipPipPipPipPipPip
  • 1,890 posts
1
Neutral

Posted 26 January 2001 - 10:00 PM

What protocol are you using? If you have enough money to buy a kit, I very highly recommend the Qiagen RNA/DNA maxi kit. It lets you extract whole RNA and Genomic DNA using the same protocol. Additionally, it lets you extract both small and large quantities of these nucleic acids (depending on how much tissue you want to use) I have used this kit many times. I have also screwed up many times (forgot to use B-mercaptoethanol once, accidentally added elution buffer at the wrong step, etc)- But even when I think I have screwed it up, I've gotten RNA with a 260/280 from 1.9-2.1 every time, usually right at 2.0.

#3 anonymous

anonymous

    Veteran

  • PipPipPipPipPipPipPipPipPipPip
  • 1,890 posts
1
Neutral

Posted 02 February 2001 - 10:00 PM

Tell me more. What's your source of message, what's the method.

Regards

Gregory


#4 anonymous

anonymous

    Veteran

  • PipPipPipPipPipPipPipPipPipPip
  • 1,890 posts
1
Neutral

Posted 25 October 2001 - 09:00 PM

i would do an extra phenol/chloroform extraction followed by just a chloroform extraction. the chloroform extract. will remove any residual phenol which will alter the OD ratio. this is of course assuming thatyou followed the historical RNA isolation method as cited by Maniatis. followed up by ppt. overnight in the -20C

#5 pon103

pon103

    member

  • Members
  • Pip
  • 3 posts
0
Neutral

Posted 30 March 2005 - 01:01 AM

Hi everyone,
I'd like to ask you if it is possible to overcome DNA contamination during RNA isolation. I use the TRIzol method and I always have DNA contamination.

Thanks in advance

#6 fred_33

fred_33

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 288 posts
0
Neutral

Posted 30 March 2005 - 02:31 AM

hi
for qiagen kit, does it allows isolation of siRNA too? i'm intersted on that point...

On the trizol manual, it's told that low ratio 260/280 may be due to a bad resuspension of rna. Try increase the amount of DEPC water of our prep. Will be better. One other possibility is to heat your rna 10' at 65 before make a OD measure.

To Pon103 :
try increase the amount of trizol you use for cell/tissue lysis. It will reduce contamination possibilities...

Fred




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

©1999-2013 Protocol Online, All rights reserved.