Help-Lg-scale Bacterial Genomic DNA Prep

Joel Bader baderja at aub.mindspring.com
Fri Aug 23 07:18:11 EST 1996


I'm seeking some advice on large-scale genomic DNA preps.  I have a
bacteria (gram-negative) that I want to pop and extract pure genomic
DNA, but the organism produces tremendous amounts of polysaccarides,
gycoproteins and other general protein contaminates.  I have up to now
been using several large scale chemical extraction methods, i.e.
Marmur procedure and Cetyltrimethylammonium Bromide lysis with little
success.  Can someone help?  I don't get any DNA in my last extraction
steps and the DNA refuses to come out of solution during the proper
ethonal extractions.  I suspect that my original lysate was
incompletely lysed or that the DNA stuck to the
polysaccarides/proteins and was spun out with with them in an earlier
phenal-CH3 extraction/centrifugation step. Thanks in advice for any
help anyone wishes to provide.
			
		Joel Bader<jabader at mindspring.com>




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