In vitro transcriptions

Matthew Thomas thomas at molbio.uoregon.edu
Wed Sep 1 19:37:03 EST 1993


In article <262067$9l6 at violet.csv.warwick.ac.uk>, lsren at csv.warwick.ac.uk
(Mr D J Plows) wrote:
> 
> 
> 		Could someone please tell me what I'm doing wrong. When
> I run aliquots of my in vitro transcription reaction of a agarose gel, I can
> see bands corresponding to the synthesised RNA. However, after 
> phenol/chloroforom extraction and ethanol precipitation, the RNA appears
>  as a smear on the agarose gel. What's going on or is this normal?
> 
> 		Thanks in advance
> 
> 		Dav

Maybe it is just salt left over from your precipitation.  Try a 70% EToh
wash
and make sure to dry the pellets.  Why are you doing this on an agarose gel
acrylamide usually looks better in my experience.  If you have a long 
transcript just use a low percentage of acrylamide (4% is the lowest I
would
go) and run it longer.

Any creative spelling is a product of my genes, and should be ignored as
much as possible!  :)

Thomas at molbio.uoregon.edu

     



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