poly(A) RNA isolation, question?

[Hiranya Roychowdhury]

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
>        Fred Hutchinson Cancer Center
>         E-mail address: stzung at fred.fhcrc.org
> 
> 

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. For a complete step-by-step 
protocol, consult any molecular biology manual - the basic principle and 
the steps are same in all the cases, following a clean total RNA prep. 
One may have to standardize the conditions, like temp., buffer conc., 
whether to include SDS etc., depending on the source organism and the 
abundance of the message of interest etc., but that should not take more 
than a a couple of days. 

			>>>>>>>>>>>>>>>>>>>>>>>>>>>
			  Hiranya S. Roychowdhury
   			  Plant Genetic Engineering Lab.
			  Box 3GL, NM State Univ.
			  Las Cruces, NM 88003
			  Phone: (505) 646-5785
			  hroychow at nmsu.edu
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