种酵母基因组提取方法的比较.PDFVIP

170 2011, Vol. 32, No. 09 食品科学 ※生物工程 4 种酵母基因组提取方法的比较 1,2 1,2 1 1 1,2 , 赵宏宇 ，李 珺 ，赵 玥 ，王 馨 ，蔡 禄 * ( 1. 内蒙古科技大学数理与生物工程学院，内蒙古包头 0 140 10 ； 2.内蒙古科技大学生物工程与技术研究所，内蒙古包头 0 140 10) 摘 要：为获得简便高效的提取酵母基因组DNA 的方法，分别采用溶菌酶法、蜗牛酶过夜处理法、蜗牛酶反复冻 融法和珠磨法处理酵母细胞，在荧光倒置显微镜下观察、比较细胞的破壁情况，提取基因组后利用紫外分光光度 计测定样品中DNA 的浓度和纯度以及琼脂糖凝胶电泳和PCR 扩增检测基因组DNA 的质量。结果表明，除溶菌酶 法外，其他3 种方法均能提取出高质量的酵母基因组DNA 。珠磨法提取酵母基因组具有质量高、成本低、时间 短且操作简单的优点。 关键词：酵母；基因组；珠磨法；蜗牛酶；溶菌酶 Comparison of Four Methods for Genomic DNA Extraction from Yeasts 1,2 1,2 1 1 1,2, ZHA O Hong-yu ，LI Jun ，ZHA O Yue ，WAN G X in ，CAI Lu * (1. School of Mathematics, Physics and Biological Engineering, Inner Mongolia University of Science and Technology, Baotou 0 140 10, China ；2. Institute of Bioengineering and Technology, Inner Mongolia University of Science and Technology, Baotou 0 140 10, China) Abstract ：In order to screen a simple and effective method for the extraction of yeast genomic DNA, different methods, such as lysozyme digestion, overnight snailase digestion, snailase digestion followed by repeated freezing and thawing and grinding with glass beads were compared for their performance based on cell disruption observed under inverted fluorescence microscope, DNA concentration and purity determined using UV spectrophotometer and DNA quality detected by agarose gel electrophore- sis and PCR. Except lysozyme digestion, all other methods provided high-quality yeast genomic DNA. The method of grinding with glass beads presented the advantages of high quality extracts, lower cost, less time