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狮子头热泉菌席样品环境总DNA提取方法的比较研究
引用本文:任禛,王治刚,尹敏,刘继辉,李沁元,王永霞,崔晓龙.狮子头热泉菌席样品环境总DNA提取方法的比较研究[J].微生物学杂志,2009,29(5):1-7.
作者姓名:任禛  王治刚  尹敏  刘继辉  李沁元  王永霞  崔晓龙
作者单位:云南大学云南省微生物研究所,云南,昆明,650091
基金项目:国家高技术研究发展计划,国家自然科学基金,云南省自然科学基金 
摘    要:通过对狮子头热泉7个环境菌席样品所提取的总DNA进行纯度检测、提取得率计算和DGGE分析,比较了3种直接和1种间接DNA提取方法。结果表明:综合利用多种裂解方式比单一裂解方式更能充分释放环境DNA;其中3种方法获得的DNA片段能够进行后续16S rDNA扩增;针对同一样品,不同方法提取的环境DNA,可获得不同DGGE群落指纹图谱;间接提取法提取的总DNA,能更好地反映狮子头热泉菌席的微生物多样性。

关 键 词:热泉环境  微生物菌席  环境DNA  DNA提取方法

Comparison of Environmental Total DNA Extraction Methods from Microbial Mat Samples in Shizitou Hot Spring
REN Zhen,WANG Zhi-gang,YIN Min,LIU Ji-hui,LI Qin-yuan,WANG Yong-xi,CUI Xiao-long.Comparison of Environmental Total DNA Extraction Methods from Microbial Mat Samples in Shizitou Hot Spring[J].Journal of Microbiology,2009,29(5):1-7.
Authors:REN Zhen  WANG Zhi-gang  YIN Min  LIU Ji-hui  LI Qin-yuan  WANG Yong-xi  CUI Xiao-long
Institution:(Lab. of Mol. Ecol. , Yunnan Inst. of Microbiol. , Yunnan Univ. , Kunming 650091 )
Abstract:Three different direct and one indirect DNA extraction methods through 7 microbial mat samples from Shiz-itou hot spring environment and the DNA test and determination of their purity, the calculation of the recovery of the extraction and DGGE analysis were compared. The results showed that comprehensively using multiple lysis methods could much more fully release the environmental DNA than a single one; and DNA fragments obtained by the three methods could carry out the following amplification of 16S rDNA ; aiming the same samples, extraction of environmen-tal DNA with different methods could obtain different DGGE community fingerprints atlas. With indirect extraction methods to extract total DNA could better express mat microbial diversity in Shizitou hot spring.
Keywords:hot spring  microbial mat  environmental DNA  DNA extraction method
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