【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.更多还原

【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.更多还原