title: Comparison of seven methods for extraction of bacterial DNA from fecal and cecal samples of mice.
abstract: 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.
authors.affiliation: EA 7300 Stress Immunité Pathogènes, Université de Lorraine, Vandoeuvre-lès-Nancy, France. Electronic address: j.ferrand@chu-nancy.fr.
authors.forename: Janina
authors.initials: J
authors.lastname: Ferrand
authors.affiliation: EA 7300 Stress Immunité Pathogènes, Université de Lorraine, Vandoeuvre-lès-Nancy, France.
authors.forename: Kevin
authors.initials: K
authors.lastname: Patron
authors.affiliation: EA 7300 Stress Immunité Pathogènes, Université de Lorraine, Vandoeuvre-lès-Nancy, France.
authors.forename: Christine
authors.initials: C
authors.lastname: Legrand-Frossi
authors.affiliation: EA 7300 Stress Immunité Pathogènes, Université de Lorraine, Vandoeuvre-lès-Nancy, France.
authors.forename: Jean-Pol
authors.initials: JP
authors.lastname: Frippiat
authors.affiliation: Université de Lorraine-CNRS, Laboratoire de Chimie Physique et Microbiologie pour l'Environnement (LCPME), UMR, 7564 Vandœuvre-lès-Nancy, France.
authors.forename: Christophe
authors.initials: C
authors.lastname: Merlin
authors.affiliation: EA 7300 Stress Immunité Pathogènes, Université de Lorraine, Vandoeuvre-lès-Nancy, France.
authors.forename: Corentine
authors.initials: C
authors.lastname: Alauzet
authors.affiliation: EA 7300 Stress Immunité Pathogènes, Université de Lorraine, Vandoeuvre-lès-Nancy, France.
authors.forename: Alain
authors.initials: A
authors.lastname: Lozniewski
mesh_terms: D000818:Animals; D001419:Bacteria / Q000235:genetics* / Q000302:isolation & purification; D002432:Cecum / Q000382:microbiology*; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D004586:Electrophoresis; D005243:Feces / Q000382:microbiology*; D005470:Fluorometry; D008297:Male; D008810:Mice, Inbred C57BL; D008967:Molecular Biology / Q000379:methods*; D060888:Real-Time Polymerase Chain Reaction; D013048:Specimen Handling / Q000379:methods*; D013053:Spectrophotometry
chemical_list: D004269:DNA, Bacterial
keywords: 16S rDNA; DNA extraction; Mice cecal contents; Mice feces; qPCR