RNA from CsCl or Quiagen plasmid?
a.hanly at cheque.uq.oz.au
Tue Mar 29 02:26:17 EST 1994
On Sat, 26 Mar 1994 23:01:50 -0500,
Steven Coon <sdcoon at umich.edu> wrote:
>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.
>sdcoon at umich.edu
>University of Michigan Medical Center
I totally agree too.
I've used Quiagen prepared DNA for in vitri transcription without problems.
(I also add RNase with each prep and use fresh lysis solution)
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