Using Glycogen to prec DNA???
crasmussen at anat.med.ualberta.ca
Wed Nov 9 10:54:05 EST 1994
In article <39p5kb$o1c at nermal.cs.uoguelph.ca>, sannis at uoguelph.ca (Seanna
> 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)
The normal amount to use is about 20 ug per precipitation...Boehringer's
insert sheet gives a suggested value. The way we purify fragments is to
run them in low melting temp agarose, melt the gel slice and use the
Promega Wizard Clean-up kit to extract the DNA from the gel. The yields
are great and there is on precipitation required at all.
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