antisense

Wolfgang Schechinger Wolfgang.Schechinger at med.uni-tuebingen.de
Fri Mar 31 09:12:27 EST 2000


Rob, 

there are no silly questions. Not asking them is silly. Did you ever 
notice that you lost knowledge when you were asking something?

Antisense mRNA probably will be degraded as soon as you add it to the 
TC flask. You may transcribe it in to cDNA, clone it into a plasmid 
and transfect this plasmid. 
If you should want to use antisense oligos, synthesize full 
phosphorothioates. They are expensive (longer synthesis time), but 
very stable. So you'll be lucky if you know someone owning a 
synthesizer...
When you design the probes, I'd suggest to make some 
target mRNA structure calculations (Ralf Zucker's RNAFOLD is an 
internet tool for this) and place them in the loops, not in the 
stems (except you should want to stabilize your target).

There is an IMHO intersting journal on this topic, Antisense Research 
and Development.

All the best, 

Wolfgang

> From:          rh at mblab.gla.ac.uk (Robert Hartley)
> Subject:       antisense
> Date:          Fri, 31 Mar 2000 13:51:44 +0100
> Organization:  Centre for Cell Engineering
> To:            methods at hgmp.mrc.ac.uk

> hello all.
> This is a really basic and silly Q?
> We have some antisense RNA and I was wondering whether this can be
> put into media for its cellular action (RNase activity?) and if we
> can use AS DNA which is cheaper? This would let me play about a bit
> more.
> 
> PS I know a lit search and a hunt about will eventually get the info
> but I also know your collective knowlege will be better.
> 
> Ok enough of the grovelling :-)
> 
> 
> Cheers
> Bob
> 
> -- 
> Robert Hartley,
> Centre for Cell Engineering,University of Glasgow,UK.
> mail: rh at mblab.gla.ac.uk, Tel: ++44 (0)141 330 4756
> Web : http://www.gla.ac.uk/Inter/CellEngineering
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Dr. Wolfgang Schechinger, Dept. of Pathobiochemistry
University of Tuebingen, Germany
email: wolfgang.schechinger at med.uni-tuebingen.de 
wwWait: http://www.medizin.uni-tuebingen.de/~wgschech/start.htm
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