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《Chinese Journal of Microecology》 2008-01
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A comparison of three protocols for the extraction of total fecal DNA

SHEN Jian,ZHANG Bao-rang,WEI Hua,PANG Xiao-yan,ZHAO Li-ping(School of Life Science and Biotechnology,Shanghai Jiao Tong University,Shanghai 200240,China)  
Objective To investigate the effects of different fecal DNA extraction methods on the molecular analysis of the diversity of gut microflora.Methods Three protocols-bead beating,chemical lysis method and QIAamp DNA Stool Mini Kit were used to extract total DNA from one fecal sample of an adult.The DNA yield and the DGGE profiles of the amplicons of 16S rRNA gene V3 region of the three DNA isolation protocols were compared.Results The DNA yield of bead beating was 2 times higher than that of the other two protocols.The Dice coefficient between DGGE profiles of these three protocols was from 60%~70%.Two predominant bands only appeared in the DGGE patterns obtained with the bead beating method.The DNA isolation experiments using 2~5 min duration of bead beating showed that DNA yield slightly increased with the increasing beating time but the DGGE patterns remained constant.Conclusion Different DNA isolation methods can affect the diversity analysis of the microbiota due to their different lysing efficiency,bead beating is more appropriate for DNA extraction from fecal samples because it can give higher DNA yield and recover more diversity.
【CateGory Index】: R346
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