Zhang D, Yang Y, Castlebury L A, Cerniglia C E
Division of Microbiology, National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, AR 72079, USA.
FEMS Microbiol Lett. 1996 Dec 1;145(2):261-5. doi: 10.1111/j.1574-6968.1996.tb08587.x.
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.
报道了一种从接合菌雅致小克银汉霉以及其他丝状真菌和酵母中分离基因组DNA的方法。该方法包括使用干冰研磨来破碎细胞，用十六烷基三甲基溴化铵并通过酚抽提去除多糖，以及在室温下用异丙醇沉淀DNA。该分离方法可大规模生产（约1mg DNA/5g湿细胞）且高度纯化的高分子量DNA。当Sau3AI部分消化的DNA与ZAP-express λ载体连接时，显示出高转化效率（>10⁶/100ng DNA）。
