| 冰川表层雪样中微生物总DNA几种提取方法的比较研究 |
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| 引用本文： | 闫沛迎,侯书贵,陈拓,张淑红,孙维君.冰川表层雪样中微生物总DNA几种提取方法的比较研究[J].冰川冻土,2012,34(2):478-484. |
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| 作者姓名： | 闫沛迎 侯书贵 陈拓 张淑红 孙维君 |
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| 作者单位： | 中国科学院寒区旱区环境与工程研究所冰冻圈科学国家重点实验室,甘肃兰州,730000;中国科学院寒区旱区环境与工程研究所冰冻圈科学国家重点实验室,甘肃兰州730000／南京大学地理与海洋科学学院,江苏南京210093;商丘师范学院生命科学系,河南商丘,476000 |
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| 摘 要： | 通过对4种具有不同裂解方式的微生物总DNA提取方法的比较,筛选出较佳的方法进行滤膜处理、DNA抽提及沉淀的优化,再将优化出的传统DNA提取方法与商业化试剂盒提取方法比较.结果表明:商业化试剂盒提取总DNA的方法适用于冰川表层雪微生物总DNA的提取;其次,将滤膜剪碎,用溶菌酶(5mg.mL-1)+蛋白酶K(1mg.mL-1)+CTAB(1%)+SDS(1%)的裂解方式,氯仿:异戊醇(24∶1)抽提一次,乙醇沉淀DNA的提取方法是一种效果较佳且廉价的冰川表层雪微生物总DNA提取方法.
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| 关 键 词： | 冰川表层雪 微生物总DNA 方法优化 |
Comparison of Methods for Total DNA Extraction of Microorganisms in Glacier Surface Snow |
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| YAN Pei-ying,HOU Shu-gui,CHEN Tuo,ZHANG Shu-hong,SUN Wei-jun.Comparison of Methods for Total DNA Extraction of Microorganisms in Glacier Surface Snow[J].Journal of Glaciology and Geocryology,2012,34(2):478-484. |
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| Authors: | YAN Pei-ying HOU Shu-gui CHEN Tuo ZHANG Shu-hong SUN Wei-jun |
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| Institution: | 1(1.State Key Laboratory of Cryospheric Sciencess,Cold and Arid Regions Environmental and Engineering Research Institute,Chinese Academy of Sciences,Lanzhou Gansu 730000,China;2.School of Geographic and Oceanographic Sciences,Nanjing University,Nanjing Jiangsu 210093,China;3.Department of Life Science,Shangqiu Normal University,Shangqiu Henan 476000,China) |
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| Abstract: | A better method was screened out through comparison of the four methods for total DNA extraction of microorganisms with different lysis.The method screened out was used to optimize the experiments of treating filter membrane,DNA extraction and precipitation.Finally,the optimized method was compared with commercial kit for total DNA extraction of microorganisms.The results showed that the commercial kit was fit for total DNA extraction of microorganisms in surface snow on glacier,and another method with cutting the filter membrane into pieces,lysis(lysozyme(5 mg·mL-1) + proteaseK(1 mg·mL-1) +CTAB(1%)+SDS(1%)),extraction once by chloroform-isoamylalcohol(24∶1) and DNA precipitation by ethanol,was an effective and low-cost method for total DNA extraction of microorganisms in surface snow on glacier. |
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| Keywords: | glacier surface snow total DNA extraction of microorganisms optimizing method |
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