RNA isolation- the long and the short of it...

Shelley Cole shelley at DARWIN.SFBR.ORG
Thu Mar 28 13:38:28 EST 1996


I have noticed this as well, but do not have an explanation or solution.

Shelley


On Thu, 28 Mar 1996, przemko wrote:

> Hi!
> I have noticed something that I wonder anyone else has seen as well. When one is isolating RNA 
> using the old fashioned Cscl gradient, one get ribosomal bands (28 and 18) in about 2:1 ratio (on 
> nondenaturing agarose gel) and virtually no small stuff (5.3 etc).
> Now, using Chomczynski method (in fact TRIZOL from Gibco) I get ribosomal bands in 1:1 or even 1:2 
> ratio and a lot of the small stuff (5.3, 5.8 etc). My RNA is not degraded as judged by subsequent 
> northern.
> Now, I can imagine that during a spin (Cscl) the denser stuff comes first, so small open molecules 
> are lost. In Trizol, the big stuff seems to get underextracted (presumably partitiones to the 
> interphase?).
> Has anyone else made similar observations? Has anyone adapted the Trizol (Chomczynski) method to 
> get more long RNA out?
> 
> ThankX
> 
> Przemko
> 
> 
> przemko at sgi.celgen.kuleuven.ac.be
> 
> 



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