Simple and reliable genotyping protocol for mouse Prkdc(SCID) mutation

J Immunol Methods. 2016 Apr:431:60-2. doi: 10.1016/j.jim.2016.02.002. Epub 2016 Feb 3.

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.

Keywords: Genotyping; Humanized mice; NSG; RFLP; SCID.

Publication types

  • Research Support, N.I.H., Extramural
  • Validation Study

MeSH terms

  • Animals
  • DNA Mutational Analysis / methods*
  • DNA-Activated Protein Kinase / genetics*
  • DNA-Binding Proteins / genetics*
  • Disease Models, Animal
  • Genotyping Techniques / methods*
  • Mice
  • Mice, SCID
  • Mutation / genetics*
  • Nuclear Proteins / genetics*
  • Polymerase Chain Reaction
  • Polymorphism, Restriction Fragment Length / genetics*

Substances

  • DNA-Binding Proteins
  • Nuclear Proteins
  • DNA-Activated Protein Kinase
  • Prkdc protein, mouse