Exploratory Animal and Medical Research · Published 2026-06-01 · DOI 10.52635/eamr/16.1.28-39
Nopparat Phumkumarn, Jarasporn Tangkhabuanbutra, Piyapha Hirunpatrawong, Panya Chariyahpongpun
Single nucleotide polymorphisms (SNPs) are an effective method for determining the genetic background of ICR outbred mice in the National Laboratory Animal Center, Mahidol University. We screened 20 SNPs on all autosomes and X chromosome. This method provides fast, accurate and reliable with high confidence for assessing the genetic background of the mice used in research. The SNP markers used in this study were obtained from publicly available SNP databases of inbred mice in the ICLAS Monitoring Center and were subsequently designed using MassARRAY assay design software. Tail samples from ICR outbred mice were collected for genomic DNA extraction. We amplified gene fragments using multiplex PCR and analyzed them through allele-specific primer extension, based on matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS). We described the 20 multiplexes PCR assays for detection with the Sequenom MassARRAY® system. The genetic monitoring reference DNA samples from inbred mice were analyzed for quality control, demonstrating 100% accuracy. The MassARRAY® TYPER 4.0 genotyping software automatically determined phenotypes based on peak heights and mass. Among the analyzed SNPs, nine SNPs provided a heterozygous genotype frequency. No deviation from Hardy – Weinberg equilibrium (PHWE > 0.05) was detected for all SNPs. All nine SNPs exhibited heterozygote deficiency, as indicated by positive FIS values. These polymorphic SNPs offer in this study provide preliminary insights into genetic variability and may serve as candidate markers for future validation and refinement of genetic monitoring within outbred strains.
Abstract from DOAJ. Public domain (CC0 1.0).
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Phumkumarn, N., Tangkhabuanbutra, J., Hirunpatrawong, P., et al. (2026). SINGLE NUCLEOTIDE POLYMORPHISMS APPROACH FOR GENETIC BACKGROUND OF ICR MICE USING MASSARRAY. Exploratory Animal and Medical Research. https://doi.org/10.52635/eamr/16.1.28-39