Favorite DNA isolation protocol

Brett Tyler fztyler at ISUN.UCDAVIS.EDU
Tue Nov 7 18:26:33 EST 1995


We have used the method of Rader and Broda (Lett. Appl. Microbiol. 1:
17-20) without modification very successfully  for a wide range of fungi
over the years including Neurospora crassa, Aspergillus spp., Trichoderma
spp., Cladosporium spp., Phytophthora spp., Armillaria spp. and truffle
fungi.  The very long centrifugation appears to get rid of a lot of
carbohydrate.

Brett Tyler

>I apologize if this topic has shown up before on this news group.
>But, I was wondering what methods fellow molecular biologist
>working with filamentous fungi prefer for isolating genomic DNA
>from fungi.  I've tried numerous methods and am not very content
>with any particular one.  Do most of you use CsCl or CTAB for
>getting rid of carbohydrate?  The best luck I have had with
>isolating high quality genomic DNA has been with grinding in
>liquid nitrogen (with mortar and pestle), extracting a few times
>with phenol/chlor, and finally separating with CsCl.
>Unfortunately, this procedure is very time consuming.  Any other
>input??
>
>Christopher D. Skory, Ph.D.
>Research Microbiologist
>
>National Center for Agricultural
>  Utilization Research, USDA-ARS
>Fermentation Biochemistry Unit
>Peoria, Illinois 61571-3902
>
>E-mail:  skorycd at ncaur1.ncaur.gov


****************
Brett Tyler
Professor
Department of Plant Pathology
University of California, Davis
bmtyler at ucdavis.edu
****************





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