RNA from CsCl or Quiagen plasmid?

Steven Coon sdcoon at umich.edu
Sat Mar 26 23:01:50 EST 1994


In article <2mqhle$40d at mserv1.dl.ac.uk>, "JOHNSON_T"
<johnson_t at am.pclsv3.sandoz.com> wrote:

> I remember looking at a few articles on the network 
> regarding RNA transcription from plasmids, but at the 
> time I discarded them because I simply could not read all 
> the postings.  I now would like the information but I 
> cannot access the archives from my computer.  Company 
> security only allows us to send and receive mail.  I 
> would greatly appreciate it if some one could tell me 
> about RNA transcription from Quiagen/Wizard plasmid preps 
> vs. CsCl.  Does the RNase used in the Quiagen/Wizard 
> preparations destroy in vitro transcripts.  I suppose it 
> would be better to use CsCl purified plasmid instead.
 

I totally agree with the other posting. The Qiagen columns work very well
and much better than CsCl. However note that the lysing solution eventually
goes bad and so the yeild declines with time. But all you need to do is
make up new stuff which consists of 0.2M NaOH,1% SDS in water. The columns
will give you 2-5 times more yeild in general and it can all be done in one
afternoon. Contamination of genomic DNA is almost zero. The kit provides
you with RNase but I always add alittle fresh RNase A to solution I before
I procede anyway. Besides CsCl does not totally eliminate RNA or genomic
unless you spin several times and you have to treat those samples with
RNAse too. Alot more work for less plasmid.

Steve Coon
sdcoon at umich.edu
University of Michigan Medical Center



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