摘要
目的:为了从分子水平上了解厌氧颗粒污泥中微生物的种类和数量,研究一种高效提取环境微生物DNA的方法。方法:厌氧颗粒污泥样品经液氮速冻、沸水浴融化、溶菌酶处理和SDS裂解后,琼脂糖凝胶电泳检测所提取的DNA,以提取的总DNA为模板,进行细菌核糖体小亚基16S rDNA基因V8、V9区的PCR扩增。结果:经检测,其DNA片段约为20 kb,样品D260nm/D280nm值为1.88,扩增结果理想,与OMEGA公司提供的试剂盒提取效果基本一致。结论:为薯类酒糟厌氧发酵污泥中微生物群落的分子生态学研究提供了一种简便、可靠的DNA提取方法。
Objective： In order to research the variety and quantity of anaerobic granular sludge on molecular level,a rapid and less instrument-dependent protocol for direct extraction of DNA from anaerobic granular sludge was developed.Methods： The thermal shock,lysozyme and SDS were used to disrupt dispersed cells and release DNA.The quality and quantity of extracted DNA was determined by agarose gel electrophoresis.The V8 and V9 region of 16S rDNA were also amplicated to test whether the DNA could be further molecular ecology studies.Results： The results showed that the isolated DNA was about 20 kb in size and the D260nm/D280nm was over 1.8,the amplification of V8 and V9 region of 16S rDNA was successed.Conclusion： This novel DNA extraction method was suitable for molecular microbial ecology studies in granular sludge system.
出处
《生物技术通讯》
CAS
2012年第1期112-114,共3页
Letters in Biotechnology
基金
国家科技支撑计划(2007BAD41B00)
关键词
颗粒厌氧污泥
DNA提取
PCR扩增
anaerobic granular sludge
DNA extraction
PCR amplification
