RNA from CsCl or Quiagen plasmid?

Andrew Simmonds asimmond at gpu.srv.ualberta.ca
Wed Mar 23 22:04:53 EST 1994


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.

: Thanks in advance

: *********************************************************
: Todd M. Johnson
: Sandoz Research Institute

: E-mail  Johnson_T at AM.pclsv3.sandoz.com

I make RNA transcripts from plasmids often and I NEVER have had to use 
CsCl purified plasmids.  I find a simple alkaline lysis (employing a 
phenol/chloroform step after RNAse or alternatively using the Quiagen 
column) works quite well.  I then cut my plasmids and run out my linearized 
templates on an agarose gel and re-isolate them from that.  This gives me 
oodles of RNA and no degredation as far as I can see on RNA gels.
Andrew
______________________________________________________________________________
Andrew Simmonds - Department of Genetics - University of Alberta CANADA
ASIMMOND at GPU.SRV.UALBERTA.CA
ASIMMOND at VM.UCS.UALBERTA.CA
G216 Biological Sciences Centre
Edmonton, Alberta, CANADA T6G 2E9
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