一种从活性污泥中提取微生物总DNA的方法  

The Extraction Method of Bacterial DNA from Activated Sludges

作　　者：郑维[1] 权春善[1] 朴永哲[1] 王军华[1] 范圣第[1] 

机构地区：[1]大连民族学院生物技术与资源利用国家民委-教育部重点实验室,大连116600

出　　处：《中国生物工程杂志》2008年第7期97-99,共3页China Biotechnology

基　　金：辽宁省自然科学基金(202001);大连民族学院青年基金(2007A101)资助项目

摘　　要：对活性污泥的微生物群落进行研究的首要前提是获得大量的高纯度微生物基因组DNA。建立了一种高效、简便的提取活性污泥总DNA方法。从提取的核酸总量、纯度、基因组完整性等多方面对所得到的DNA质量进行了评价,结果表明,从单位活性污泥中提取的DNA得率为105～823μg/g,结构完整,纯度很高,无需进一步的纯化,可直接进行微生物群落分析及构建文库等后续分子生物学操作。现在实验室使用的提取活性污泥中DNA的方法,纯度普遍都无法达到PCR反应和建立文库的要求,建立的活性污泥DNA提取方法则可以克服这一难题。Methods for studying the population diversity of microorganism in activated sludge usually require enrichment of bacterial genome. The efficient information on microbial species composition provided and shifted in diversity revealed are dependent on the effective DNA recovery technique. The method was based on washing by alkaline phosphate buffer and digestion with extended heating of the activated sludge suspension in the presence of lysozyme and freeze-thawing in high-salt-SDS buffer. The extraction was tested for four activated sludge differing in places and dates. The DNA fragment from all sludge was integrity. DNA yields ranged from 105 to 823μg/g sludge and were of sufficient purity for PCR-based 16S ribosomal DNA analysis and restriction digested. In general, all methods produced DNA pure were not enough for PCR amplification and libraries construction. As basis of experimental goals, the study provides an appropriate extraction method of microbial DNA in sludge.

关 键 词：活性污泥 DNA提取 PEG 16S RDNA 

分 类 号：Q523[生物学—生物化学]