Comparative Evaluation of DNA Extraction Methods from Feces of Multiple Host Species for Downstream Next-Generation Sequencing

被引:100
作者
Hart, Marcia L. [1 ]
Meyer, Alexandra [2 ]
Johnson, Philip J. [3 ]
Ericsson, Aaron C. [1 ,4 ,5 ]
机构
[1] Univ Missouri, Dept Vet Pathobiol, Comparat Med Program, Columbia, MO 65211 USA
[2] Univ Missouri, Coll Vet Med, Columbia, MO USA
[3] Univ Missouri, Dept Vet Med & Surg, Columbia, MO USA
[4] Univ Missouri, Metagen Ctr, Columbia, MO USA
[5] Univ Missouri, Mutant Mouse Resource & Res Ctr, Columbia, MO USA
来源
PLOS ONE | 2015年 / 10卷 / 11期
基金
美国国家卫生研究院;
关键词
GUT MICROBIOTA; BACTERIAL-DNA; DATABASE;
D O I
10.1371/journal.pone.0143334
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
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.
引用
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页数:16
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