A method for the large scale isolation of high transformation efficiency fungal genomic DNA.
A procedure for isolation of genomic DNA from the zygomycete Cunninghamella elegans and other filamentous fungi and yeasts is reported. This procedure involves disruption of cells by grinding using dry ice, removal of polysaccharides using cetyltrimethylammonium bromide and by phenol extractions, and precipitation of DNA with isopropanol at room temperature. The isolation method produced large scale (approximate 1 mg DNA/5 g wet cells) and highly purified high molecular mass DNA. Sau3AI partially digested DNA showed high transformation efficiency (> 10(6)/100 ng DNA) when ligated to ZAP-express lambda vector.
Zhang
D
D
Division of Microbiology, National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, AR 72079, USA.
Yang
Y
Y
Castlebury
L A
LA
Cerniglia
C E
CE
D001487:Basidiomycota / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics* / Q000302:isolation & purification; D005658:Fungi / Q000235:genetics*; D005670:Fusarium / Q000235:genetics; D015723:Gene Library; D016678:Genome; D014170:Transformation, Genetic; D014250:Trichosporon / Q000235:genetics
D004271:DNA, Fungal