best polyA+ from total RNA method??

Gary Krause gkrause at cms.cc.wayne.edu
Fri Feb 10 10:17:32 EST 1995


In article <Pine.DYN.3.91.950209140740.15540A-100000 at uxa.cso.uiuc.edu> hodges bradley l <segdoh at uxa.cso.uiuc.edu> writes:
>From: hodges bradley l <segdoh at uxa.cso.uiuc.edu>
>Subject: Re: best polyA+ from total RNA method??
>Date: Thu, 9 Feb 1995 14:09:10 -0600

>Qiagen sells dT conjugated glass beads (Ithink).
> 

>On 1 Feb 1995 neale at mbcf.stjude.org wrote:

>> I was wondering if anyone had a strong recommendation of method for
>> purification of poly A+ RNA from total RNA. The samples of total RNA that we
>> have are about 300 ug each. We could pool some if necessary.
>> 
>> We could purify poly A+ directly from cells if that is thought superior to
>> purifying from total RNA. It's just that we already have the total RNA, and
>> know that our gene of interest is expressed in these samples.
>> 
>> We wish to use the poly A+ RNA for cDNA library construction. It's been a while
>> (5 years) since I've had to do this and so I thought there may be an improved
>> method for selection of poly A+ other than oligo dT column chromatography (eg
>> magnetic beads-dT)
>> 
>>                Thanks in advance for your help.
>> 
>> Geoff Neale
>> 
>> Dept. of Virology and Molecular Biology        E-mail: neale at mbcf.stjude.org
>> St. Jude Children's Research Hospital           Phone: (901) 522-0400
>> Memphis, TN                                       Fax: (901) 523-2622
>> 
>> 
>> 
>> 



We have used Pharmacia's oligo dT spin column kit with very good results.  Try 
them.  

Gary

-------------------------------------------
Gary Krause (gkrause at cms.cc.wayne.edu)
Department of Emergency Medicine
Wayne State University, Detroit, MI USA



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