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

Rolen M Quadros, Larisa Y Poluektova, Channabasavaiah B Gurumurthy

Journal of immunological methods 2016 Apr

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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. Copyright © 2016 Elsevier B.V. All rights reserved.

Citation

Rolen M Quadros, Larisa Y Poluektova, Channabasavaiah B Gurumurthy. Simple and reliable genotyping protocol for mouse Prkdc(SCID) mutation. Journal of immunological methods. 2016 Apr;431:60-2