Optimization Method for the Extraction of Total DNA from the Intestinal Microbes in Quails

doi:10.13560/j.cnki.biotech.bull.1985.2015.01.011

Abstract

Abstract: Comparing the effects of total DNA extracted by different methods from intestinal microbes in quails, the study aimed at investigating the influences of DNA quality and PCR-DGGE conditions on intestinal microbial diversity of quail. In this study, the experiment used four methods to extract the DNA. The first was a conventional method for using saturated phenol/chloroform, other three commercially available DNA extraction kits（QIAGEN, SHENGGONG, TIANGEN）were assessed. The ribosomal gene sequences were amplified with V3 and V6-V8 hypervariable region universal primers at different annealing temperature and subjected to denaturing gradient gel electrophoresis（DGGE）. The QIAGEN DNA Kit generated the greatest bacterial species eveness and DNA quantity and quality in compare with other methods（A₂₆₀/A₂₈₀=1.8, 200 ng/μL）. The resulting DGGE profiles were substantially different in terms of the number and relative intensity of the amplification products. The V6-V8 region produced better DGGE profiles than V3 region. The QIAGEN DNA Kit and V6-V8 hypervariable region universal primers will be used in future studies of intestinal microbial diversity of quail.

Key words: quail, ntestinal microbes, DNA extraction, DGGE

Zhang Zhenhua, Bao Wangbo, Yu Ran, Wang Changyong, Liu Yan. Optimization Method for the Extraction of Total DNA from the Intestinal Microbes in Quails[J]. Biotechnology Bulletin, 2015, 31(1): 73-78.

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