Comparison of seven methods for extraction of bacterial DNA from fecal and cecal samples of mice

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...

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Bibliographic Details
Published in:Journal of microbiological methods 2014-10, Vol.105, p.180-185
Main Authors: Ferrand, Janina, Patron, Kevin, Legrand-Frossi, Christine, Frippiat, Jean-Pol, Merlin, Christophe, Alauzet, Corentine, Lozniewski, Alain
Format: Article
Language:English
Subjects:
16S rDNA
Animals
Bacteria
Bacteria - genetics
Bacteria - isolation & purification
Bacteriology
Cecum - microbiology
DNA extraction
DNA, Bacterial - genetics
DNA, Bacterial - isolation & purification
Electrophoresis
Feces - microbiology
Fluorometry
Life Sciences
Male
Mice cecal contents
Mice feces
Mice, Inbred C57BL
Microbiology and Parasitology
Molecular Biology - methods
qPCR
Real-Time Polymerase Chain Reaction
Specimen Handling - methods
Spectrophotometry
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Summary: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™ Gram Positive DNA Purification Kit, QIAamp® DNA Stool Mini Kit; NucliSENS® easyMAG®, ZR Fecal DNA MiniPrep™, FastDNA® SPIN Kit for Feces and FastDNA® 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® SPIN Kit for Soil. •Seven DNA extraction methods were compared on mice fecal and cecal samples.•DNA yield, quality and cell lysis efficiencies were evaluated.•FastDNA® SPIN Kit for Soil was the most efficient extraction method.•FastDNA® SPIN Kit for Soil was equally efficient for fecal and cecal samples.
ISSN:0167-7012
1872-8359
DOI:10.1016/j.mimet.2014.07.029