A method for the large scale isolation of high transformation efficiency fungal genomic DNA.
A method for the large scale isolation of high transformation efficiency fungal genomic DNA.
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DOI：
10.1111/j.1574-6968.1996.tb08587.x
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发表时间：
1996-12
影响因子：
2.1
通讯作者：
Donglu Zhang;Yifan Yang;L. Castlebury;C. Cerniglia
中科院分区：
文献类型：
--
作者：
Donglu Zhang;Yifan Yang;L. Castlebury;C. Cerniglia
报道了一种从放线菌Cunninghamella elegans和其它丝状真菌及酵母中提取基因组DNA的方法。该程序包括通过使用干冰研磨来破坏细胞，使用十六烷基三甲基溴化铵和通过苯酚提取来去除多糖，以及在室温下用异丙醇沉淀DNA。该分离方法产生大规模（约1 mg DNA/5g湿细胞）和高度纯化的高分子量DNA。Sau 3AI部分消化的DNA在连接到ZAP表达λ载体时显示出高转化效率（> 10（6）/100 ng 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.