title: Simple and reliable genotyping protocol for mouse Prkdc(SCID) mutation.
abstract: Mutant mouse models, genetically-engineered or spontaneous-mutations, serve as valuable tools for biomedical research. Genotyping of mutant mice is a critical requirement for maintaining the colony, to breed with other mutants and to match the phenotypic observations. The SCID (Severe Combine Immuno Deficiency) mouse model has been extensively used as a common background-strain in many immunology and transplantation studies. Many different types of assays, including Restriction Fragment Length Polymorphism (RFLP), confronting two primer pairs PCR and end-point methods have been attempted for establishing a genotyping protocol for the SCID mutation. However, the best method that is thought to be reliable is sequencing, which requires additional time and resources to perform on a routine basis. In this report, we describe a novel RFLP assay that is simple and reliable. The method is validated by sequencing analysis, and this novel method can be adapted for routine genotyping of SCID model.
authors: Mouse Genome Engineering Core Facility, Vice Chancellor for Research Office, University of Nebraska Medical Center, Omaha, NE, USA.
authors: Rolen M
authors: RM
authors: Quadros
authors: Department of Pharmacology and Experimental Neuroscience, University of Nebraska Medical Center, Omaha, NE, USA.
authors: Larisa Y
authors: LY
authors: Poluektova
authors: Mouse Genome Engineering Core Facility, Vice Chancellor for Research Office, University of Nebraska Medical Center, Omaha, NE, USA; Developmental Neuroscience, Munroe Meyer Institute for Genetics and Rehabilitation, University of Nebraska Medical Center, Omaha, NE, USA.
authors: Channabasavaiah B
authors: CB
authors: Gurumurthy
mesh_terms: D000818:Animals; D004252:DNA Mutational Analysis / Q000379:methods*; D051747:DNA-Activated Protein Kinase / Q000235:genetics*; D004268:DNA-Binding Proteins / Q000235:genetics*; D004195:Disease Models, Animal; D060005:Genotyping Techniques / Q000379:methods*; D051379:Mice; D016513:Mice, SCID; D009154:Mutation / Q000235:genetics*; D009687:Nuclear Proteins / Q000235:genetics*; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length / Q000235:genetics*
chemical_list: D004268:DNA-Binding Proteins; D009687:Nuclear Proteins; D051747:DNA-Activated Protein Kinase; C497264:Prkdc protein, mouse
keywords: Genotyping; Humanized mice; NSG; RFLP; SCID