Comparative Evaluation of DNA Extraction Methods from Feces of Multiple Host Species for Downstream Next-Generation Sequencing.
Comparative Evaluation of DNA Extraction Methods from Feces of Multiple Host Species for Downstream Next-Generation Sequencing.
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DOI：
10.1371/journal.pone.0143334
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Ericsson AC
中科院分区：
文献类型：
--
作者：
Hart ML;Meyer A;Johnson PJ;Ericsson AC
胃肠道包含大量的微生物群落，这些微生物至今仍在很大程度上无法培养，这使得该领域的研究具有挑战性。随着新的负担得起的先进测序技术，在这个令人兴奋的领域的重要突破现在是可能的。然而，样本收集、处理和DNA提取的标准化方法尚未确定。为了帮助解决这个问题，我们研究了5种常见的DNA提取方法对5种不同物种粪便样本的使用。我们的数据表明，DNA提取方法影响DNA浓度和纯度，成功的NGS扩增，并影响NGS输出中观察到的微生物群落，这取决于粪便样品的种类和使用的DNA提取方法。这些数据强调了在设计和解释跨物种研究数据时仔细考虑DNA提取方法的重要性。
The gastrointestinal tract contains a vast community of microbes that to this day remain largely unculturable, making studies in this area challenging. With the newly affordable advanced sequencing technology, important breakthroughs in this exciting field are now possible. However, standardized methods of sample collection, handling, and DNA extraction have yet to be determined. To help address this, we investigated the use of 5 common DNA extraction methods on fecal samples from 5 different species. Our data show that the method of DNA extraction impacts DNA concentration and purity, successful NGS amplification, and influences microbial communities seen in NGS output dependent on the species of fecal sample and the DNA extraction method used. These data highlight the importance of careful consideration of DNA extraction method used when designing and interpreting data from cross species studies.