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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