Comparison of seven methods for extraction of bacterial DNA from fecal and cecal samples of mice
Comparison of seven methods for extraction of bacterial DNA from fecal and cecal samples of mice
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DOI：
10.1016/j.mimet.2014.07.029
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发表时间：
2014-10-01
影响因子：
2.2
通讯作者：
Lozniewski, Alain
中科院分区：
文献类型：
--
作者：
Ferrand, Janina;Patron, Kevin;Lozniewski, Alain
从小鼠粪便样本中分析细菌DNA通常在实验研究中进行。虽然DNA提取是各种分子方法的关键步骤，但从小鼠粪便样本中提取DNA的方法的效率从未得到评估。我们比较了六种广泛使用的商用试剂盒(MasterPure （TM）革兰氏阳性DNA纯化试剂盒，QIAamp (R) DNA粪便迷你试剂盒；NucliSENS (R) easyMAG (R), ZR Fecal DNA MiniPrep (TM), FastDNA (R) SPIN Kit（粪便）和FastDNA (R) SPIN Kit（土壤），以及从小鼠粪便和盲肠内容物中分离DNA的非商业方法。采用荧光法、分光光度法、凝胶电泳法和qPCR检测DNA的数量和质量。采用qPCR方法对加标标本中3种相关细菌进行细胞裂解效率评估。对于粪便和肠道内容物，最有效的提取方法是FastDNA (R) SPIN Kit For Soil。(C) 2014 Elsevier B.V.版权所有
Analysis of bacterial DNA from fecal samples of mice is commonly performed in experimental studies. Although DNA extraction is a critical step in various molecular approaches, the efficiency of methods that may be used for DNA extraction from mice fecal samples has never been evaluated. We compared the efficiencies of six widely used commercial kits (MasterPure (TM) Gram Positive DNA Purification Kit, QIAamp (R) DNA Stool Mini Kit; NucliSENS (R) easyMAG (R), ZR Fecal DNA MiniPrep (TM), FastDNA (R) SPIN Kit for Feces and FastDNA (R) SPIN Kit for Soil) and a non-commercial method for DNA isolation from mice feces and cecal contents. DNA quantity and quality were assessed by fluorometry, spectrophotometry, gel electrophoresis and qPCR. Cell lysis efficiencies were evaluated by qPCR targeting three relevant bacteria in spiked specimens. For both feces and intestinal contents, the most efficient extraction method was the FastDNA (R) SPIN Kit for Soil. (C) 2014 Elsevier B.V. All rights reserved.