poly(A) RNA isolation, question?
Sprankle at ciit.org
Wed Dec 6 21:26:46 EST 1995
In article <Pine.SUN.3.91.951206135604.14797A-100000 at verdi>,
hroychow at NMSU.EDU (Hiranya Roychowdhury) wrote:
> On 6 Dec 1995, Shei-Pon Tzung wrote:
> > Hi all:
> > What is the best commercially available poly(A) RNA isolation kit?
> > I have tried Promega and Qiagen, which gave me a yield about 0.2 to 0.6
> > %? I have been struggling with this for months, Please help.
> > Thanks.
> > Shie-Pon Tzung
> Moving out of the 'kit-based' mindset would be a first step, I believe.
> The good old oligo-dT cellulose (or sepharose, although I prefer the
> former), in a 1-2 mL column made out of sterile 3 mL syringe and glass-wool
> has always worked with excellent recovery.
I agree. I've been very happy with bulk oligo(d)T cellulose from GIBCO/BRL,
and Bio-Rad polypropylene 12 ml columns (can't remember the exact product
name) work very well for me. I tried spin columns from various sources
several times and was never happy with them; the amount of handling
involved in using them seems to provide opportunities for contamination,
leaving you with poor quality RNA.
e-mail: sprankle at ciit.org
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