环境工程学报2012，Vol.6Issue(3)：1000-1004,5.
一种活性污泥总DNA提取方法的优化
Optimization of DNA extraction from active sludge
摘要
Abstract
Three DNA extraction methods from active sludge,classical proteinase K-SDS-chloroform/isoamyl alcohol method（CPSCI method）,liquid nitrogen skiving method and humic acids removing method were compared.CPSCI method was optimized on the quantity of sludge,incubation time,centrifugation time and precipitation time in isopropanol,respectively.The results showed that 23.1 kb genic DNA（OD260nm/OD280nm=1.86） can be obtained by optimized CPSCI method（OPSCI） under the conditions of using about 0.10g sludge,incubating 10 min at 37 ℃,shaking 5min with SDS at room temperature and centrifugating directly after adding isopropanol.The acquired DNA dilution（10 times） can be used for PCR.The OPSCI method introduced in this paper possessed good repeatability,simple operation,and high quality and yield.It was a rapid and advanced method for extracting DNA from active sludge samples in general laboratory.关键词
活性污泥/DNA提取/16S/rDNA/PCR/OPSCIKey words
active sludge/DNA extraction/16S rDNA PCR/OPSCI分类
资源环境引用本文复制引用
闫建芳,刘秋,张璐,齐小辉,张黎..一种活性污泥总DNA提取方法的优化[J].环境工程学报,2012,6(3):1000-1004,5.基金项目
国家自然科学基金资助项目 （）
中央高校基本科研业务费专项资金资助项目 （）
