Difficulties Obtaining Genomic DNA
melansond at am.abru.cg.com
Tue Dec 3 08:54:26 EST 1996
***I'm seeking protocols for genomic DNA extraction from "difficult"
***sources. Thus far, standard Proteinase K methods (published in Current
***Protocols and Maniatis) have failed miserably on the frozen tissues that
***I'm using. Commercial products, such as Bio 101's Genome kit and BRL's
***DNAzol reagent work, albeit with limited success - low yield and DNA on
***the order of 23 kb.
***One explanation that I've been entertaining recently is that I'm being
***hampered by extracellular matrix material. Is it possible to get cells
***apart with something like collagenase and still be able to inhibit
***endogeneous intracellular DNAses?
***Thanks in advance
***schoenfeld at med.utah.edu
Hi, I realize you are probably working with animal cells, but how about
good old tissue grinding under liquid Nitrogen with a mortar and pestle. If it
works for plant tissues (woody, polysaccharides, cellulose) it will probably
work for you.
More information about the Methods