摘要
基因组DNA的提取是最基本的分子生物学实验技术,介绍了一种适用于绿僵菌克隆筛选的DNA批量快速提取方法.研究结果表明:使用改良的察氏琼脂培养基(CDA)代替常用的土豆葡萄糖琼脂(PDA)培养绿僵菌,以钢珠振打破碎菌丝,配以简化的酚氯仿抽提操作,可以高效地提取高质量的真菌基因组DNA.本方法中,CDA菌落生长需2~3 d,双样品提取耗时约15 min,每个菌落可获得DNA数百纳克,片段长度在10 kb左右,电泳条带清晰可见.
Genomic DNA extraction is one of the most basic experimental techniques in molecular biology. A rapid batch DNA extraction method for the clone screening of Metarhizium was introduced. Our results indicated that using modified Czapek-Dox Agar(CDA) instead of common Potato Dextrose Agar(PDA), breaking the mycelia with metal beads and combining with simplified phenol-chloroform extraction manipulation could isolate high-quality genomic DNA efficiently. In this method, fungal colonies were supposed to grow on CDA for 2~3 days. It took about 15 minutes to accomplish extraction for a pair of samples. The DNA yield of one colony was several hundred nanograms. The fragments of DNA were around 10 kb, showing a clear band inagarose gel electrophoresis.
作者
宋红生
邓博严
殷颖
SONG Hongsheng;DENG Boyan;YIN Ying(College of Life Sciences,Shanghai University,Shanghai 200444,China;Shanghai Institute of Plant Physiology and Ecology,Chinese Academy of Sciences,Shanghai 200032,China)
出处
《信阳师范学院学报（自然科学版）》
CAS
北大核心
2020年第1期149-155,共7页
Journal of Xinyang Normal University(Natural Science Edition)
基金
国家自然科学基金项目(31501699)
关键词
机械珠磨
酚氯仿抽提
DNA分离纯化
克隆筛选
凝胶电泳分析
mechanical bead milling
phenol-chloroform extraction
dna-separation and purification
clone screening
gel-electrophoresis analysis
