Evaluation of Methods for the Extraction of DNA from Drinking Water Distribution System Biofilms

被引:44
作者
Hwang, Chiachi [1 ]
Ling, Fangqiong [1 ]
Andersen, Gary L. [2 ]
LeChevallier, Mark W. [3 ]
Liu, Wen-Tso [1 ]
机构
[1] Univ Illinois, Dept Civil & Environm Engn, Urbana, IL 61810 USA
[2] Lawrence Berkeley Natl Lab, Dept Ecol, Div Earth Sci, Berkeley, CA 94720 USA
[3] Amer Water, Voorhees, NJ 08043 USA
关键词
DNA extraction; drinking water distribution system; biofilm microbial community; POLYMERASE-CHAIN-REACTION; BACTERIAL COMMUNITIES; SOIL; PCR; PURIFICATION; IDENTIFICATION; DIVERSITY; PATHOGENS; ABUNDANCE; RICHNESS;
D O I
10.1264/jsme2.ME11132
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
While drinking water biofilms have been characterized in various drinking water distribution systems (DWDS), little is known about the impact of different DNA extraction methods on the subsequent analysis of microbial communities in drinking water biofilms. Since different DNA extraction methods have been shown to affect the outcome of microbial community analysis in other environments, it is necessary to select a DNA extraction method prior to the application of molecular tools to characterize the complex microbial ecology of the DWDS. This study compared the quantity and quality of DNA yields from selected DWDS bacteria with different cell wall properties using five widely used DNA extraction methods. These were further selected and evaluated for their efficiency and reproducibility of DNA extraction from DWDS samples. Terminal restriction fragment length analysis and the 454 pyrosequencing technique were used to interpret the differences in microbial community structure and composition, respectively, from extracted DNA. Such assessments serve as a concrete step towards the determination of an optimal DNA extraction method for drinking water biofilms, which can then provide a reliable comparison of the meta-analysis results obtained in different laboratories.
引用
收藏
页码:9 / 18
页数:10
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