title: A highly efficient strategy to determine genotypes of genetically-engineered mice using genomic DNA purified from hair roots.
abstract: Genotyping of genetically-engineered mice is necessary for the effective design of breeding strategies and identification of mutant mice. This process relies on the identification of DNA markers introduced into genomic sequences of mice, a task usually performed using the polymerase chain reaction (PCR). Clearly, the limiting step in genotyping is isolating pure genomic DNA. Isolation of mouse DNA for genotyping typically involves painful procedures such as tail snip, digit removal, or ear punch. Although the harvesting of hair has previously been proposed as a source of genomic DNA, there has been a perceived complication and reluctance to use this non-painful technique because of low DNA yields and fear of contamination. In this study we developed a simple, economic, and efficient strategy using Chelex® resins to purify genomic DNA from hair roots of mice which are suitable for genotyping. Upon comparison with standard DNA purification methods using a commercially available kit, we demonstrate that Chelex® efficiently and consistently purifies high-quality DNA from hair roots, minimizing pain, shortening time and reducing costs associated with the determination of accurate genotypes. Therefore, the use of hair roots combined with Chelex® is a reliable and more humane alternative for DNA genotyping.
authors.affiliation: 1 Department of Pharmacology & Toxicology, Wright State University, Boonshoft School of Medicine, Dayton, OH, USA.
authors.forename: Víctor
authors.initials: V
authors.lastname: Otaño-Rivera
authors.affiliation: 1 Department of Pharmacology & Toxicology, Wright State University, Boonshoft School of Medicine, Dayton, OH, USA.
authors.forename: Amma
authors.initials: A
authors.lastname: Boakye
authors.affiliation: 1 Department of Pharmacology & Toxicology, Wright State University, Boonshoft School of Medicine, Dayton, OH, USA.
authors.forename: Nadja
authors.initials: N
authors.lastname: Grobe
authors.affiliation: 1 Department of Pharmacology & Toxicology, Wright State University, Boonshoft School of Medicine, Dayton, OH, USA.
authors.forename: Mohammed M
authors.initials: MM
authors.lastname: Almutairi
authors.affiliation: 1 Department of Pharmacology & Toxicology, Wright State University, Boonshoft School of Medicine, Dayton, OH, USA.
authors.forename: Shams
authors.initials: S
authors.lastname: Kursan
authors.affiliation: 1 Department of Pharmacology & Toxicology, Wright State University, Boonshoft School of Medicine, Dayton, OH, USA.
authors.forename: Lesan K
authors.initials: LK
authors.lastname: Mattis
authors.affiliation: 2 Institute of Physiology, University of Regensburg, Regensburg, Germany.
authors.forename: Hayo
authors.initials: H
authors.lastname: Castrop
authors.affiliation: 3 Division of Nephrology, Department of Medicine, Duke O'Brien Center for Kidney Research, Duke University, Durham, NC, USA.
authors.forename: Susan B
authors.initials: SB
authors.lastname: Gurley
authors.affiliation: 1 Department of Pharmacology & Toxicology, Wright State University, Boonshoft School of Medicine, Dayton, OH, USA.
authors.forename: Khalid M
authors.initials: KM
authors.lastname: Elased
authors.affiliation: 4 Department of Pathology, Wright State University, Boonshoft School of Medicine, Dayton, OH, USA.|5 Veterans Affairs Medical Center, Cincinnati, OH, USA.
authors.forename: Gregory P
authors.initials: GP
authors.lastname: Boivin
authors.affiliation: 1 Department of Pharmacology & Toxicology, Wright State University, Boonshoft School of Medicine, Dayton, OH, USA.
authors.forename: Mauricio
authors.initials: M
authors.lastname: Di Fulvio
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics; D002614:Chelating Agents / Q000737:chemistry*; D004247:DNA / Q000302:isolation & purification*; D005260:Female; D060005:Genotyping Techniques / Q000379:methods*; D018859:Hair Follicle / Q000737:chemistry*; D008297:Male; D051379:Mice / Q000235:genetics*; D011137:Polystyrenes / Q000737:chemistry*; D011145:Polyvinyls / Q000737:chemistry*
chemical_list: D002614:Chelating Agents; D011137:Polystyrenes; D011145:Polyvinyls; C006960:chelex; D004247:DNA
keywords: 3Rs; DNA isolation; alternatives; rodents; welfare