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传统的微生物分离与培养技术无法揭示微生物群落的动态变化.为了阐释虾池沉积环境微生态格局的原始组成情况,本研究先以TENP缓冲液去除沉积物中的腐殖酸,继之以溶菌酶-SDS温和裂解,其总DNA的提取效率达90%以上,所获DNA产量达2~20μg/g沉积物(湿),片段大小均在23 kb左右,不经纯化即可直接进行PCR扩增和限制性酶切.以该DNA为模板进行PCR-DGGE分析,揭示了虾池沉积物丰富的微生物多样性.该方法是一种适用于虾池沉积物总DNA提取的简便、可靠方法.
The traditional microbial isolation and culture technology can not reveal the dynamic changes of microbial community.In order to explain the original composition of the ecological environment of the shrimp pond sediment environment, TENP buffer was used to remove the humic acid in sediments, followed by lysozyme -SDS, the total DNA extraction efficiency was more than 90%, and the DNA yield was 2 ~ 20μg / g sediment (wet), the fragment size was about 23 kb, which could be directly amplified without PCR And restriction enzyme digestion.The PCR-DGGE analysis using this DNA as a template revealed the abundance of micro-organisms in shrimp pond sediments, a simple and reliable method suitable for total DNA extraction from shrimp pond sediments.