How to clean DNA which may have glycosaminoglycans
Warren Gallin
wgallin at gpu.srv.ualberta.ca
Tue Aug 19 17:56:01 EST 1997
In Article <33FA1F8E.655D at Bristol.ac.Uk>, Harry Witchel
<Harry.Witchel at Bristol.ac.Uk> wrote:
>Good day Molecular Charmers!
> I have a problem with my DNA. Boy do I have a problem! My DNA
>comes from a marine invertebrate which is hypocellular and filled with
>gelatinous mesoglea which I'm told is made up of glycosaminoglycans or
>hyaluronic acid. This thing is mostly connective tissue.
> I have made the DNA, the animals are long gone, I have a limited
>quantity. I have phenoled, chloroformed, ethanoled and isopropanoled
>till blue in the face, but still, the stuff does not digest well with
>restriction endonucleases. Even after running down the purified DNA
>through Qiagen's genomic column, this DNA does not want to cut to
>completion.
The two methods that have worked for me are 1) CTAB purification and 2)
Chelex purification. I believe that you will find both of these in the
Current Protocols in Molecualr Biology book, probably in other manuals. If
not, drop me a line and I'll dig out my files and find a procedure for you.
Warren Gallin
Department of Biological Sciences
University of Alberta
Edmonton, Alberta T6G 2E9
Canada
wgallin at gpu.srv.ualberta.ca
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