Using Glycogen to prec DNA???
mbxspd at unicorn.nott.ac.uk
Wed Nov 9 07:51:23 EST 1994
> I am trying to purify a PCR fragments, around 600 bp from gel. I can get
> a decent amount out of the gel, but I lose it during the precipitation
> step. I want to use this product for direct cycle sequencing, so yeast
> tRNA as an aid to precipitating the DNA was suggested to me to be a bad idea.
> I have heard of glycogen used to prec oligonucleotides. Does anyone know
> of the right concentration of glycogen to use. And does it interfer with
> sequencing afterwards? Or does anyone know of another method to get a
> better yield.
> Thanks. Seanna (sannis at uoguelph.ca)
I use glycogen on a regular basis for precipitating small
fragments of DNA for all sorts of protocols. We use the RNase/DNase
free glycogen available from Boehringer-Mannheim. I think it used
to come at a conc. of 20mg/ml. I use it at a final concentration of
approx. 20 microgrammes/ml I think. Something like that. I precipitate
as normal after adding glycogen i.e with NaOAc/EtOH.
As far as it interfering with anything...I'm not sure about
sequencing but it is certainly inert in ligations, cDNA reactions in
fact I don't think we've come across a problem with it re: messing
up enzymic reactions.
Hope that helps! :)
Queens Medical Centre
Internet email: mbxspd at unicorn.nott.ac.uk
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