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首页> 外文期刊>African Journal of Microbiology Research >Comparison of DNA extraction methods for polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) analysis of the infant fecal microbial communities
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Comparison of DNA extraction methods for polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) analysis of the infant fecal microbial communities

机译：婴儿粪便微生物群落的聚合酶链反应-变性梯度凝胶电泳（PCR-DGGE）分析的DNA提取方法的比较

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The study of the intestinal microflora has been developed mainly using conventional microbiological approaches. These studies have made ??great progress, but it is imperative that new methods will be applied to facilitate scientific progress. Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) has been used to study microbial communities in many environmental samples. So, we must do our best to enhance the quality and accuracy of upstream analyses, such as DNA extraction. In this study, the relative efficacy of four DNA extraction methods (QIAamp DNA stool mini kit method, Q; QIAamp DNA stool mini kit+ Bead beating method, QB; QIAamp DNA stool mini kit+ Frozen thawed method, QF; E.Z.N.A. Stool DNA Kit method, E) were evaluated. Further, PCR-DGGE technique was also assessed in detecting diversity in infant intestinal bacterial fingerprint profiles. The total DNA was extracted from the infant fecal specimens using four different methods, followed by PCR amplification of bacterial 16S rRNA-V3 region, and DGGE separation of the amplifications. The number of extracted DNA of the infant feces using the Q method was larger than the QB, QF and E methods, and the produced bands of its DGGE profiles were more than the other three methods, which was due to its cracking temperature (95°C). We concluded that DGGE of 16S rRNA gene PCR products was suitable for capturing the profiles of the infant intestinal microbial community and the QIAamp DNA stool mini kit method was appropriate for infant fecal DNA extraction. PCR-DGGE could be an important tool for DNA studies.
机译：肠道微生物区系的研究主要是使用常规微生物学方法进行的。这些研究取得了长足的进步，但是必须应用新的方法来促进科学进步。聚合酶链反应-变性梯度凝胶电泳（PCR-DGGE）已用于研究许多环境样品中的微生物群落。因此，我们必须尽最大努力提高上游分析（例如DNA提取）的质量和准确性。在这项研究中，四种DNA提取方法的相对功效（QIAamp DNA粪便微型试剂盒方法，Q; QIAamp DNA粪便微型试剂盒+珠打法，QB; QIAamp DNA粪便微型试剂盒+冻融法，QF; EZNA凳DNA试剂盒方法， E）进行了评估。此外，还对PCR-DGGE技术进行了评估，以检测婴儿肠道细菌指纹图谱的多样性。使用四种不同方法从婴儿粪便标本中提取总DNA，然后对细菌的16S rRNA-V3区进行PCR扩增，并对扩增进行DGGE分离。采用Q法提取婴儿粪便的DNA数量要多于QB，QF和E法，其DGGE谱图产生的条带要比其他三种方法多，这是由于其裂解温度（95°C）所致。 C）。我们得出的结论是16S rRNA基因PCR产物的DGGE适用于捕获婴儿肠道微生物群落的特征，而QIAamp DNA粪便微型试剂盒方法适用于婴儿粪便DNA的提取。 PCR-DGGE可能是DNA研究的重要工具。

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