Genetic sex determination of mice by simplex PCR.
Biol Sex Differ
Springer Science and Business Media LLC
MetadataShow full item record
Tunster, S. J. (2017). Genetic sex determination of mice by simplex PCR.. Biol Sex Differ, 8 (1), 31. https://doi.org/10.1186/s13293-017-0154-6
BACKGROUND: Investigating fetal development in mice necessitates the determination of fetal sex. However, whilst the sex of adult and juvenile mice can be readily distinguished from anogenital distance, the sex of fetal and neonatal mice cannot be identified visually. Instead, genetic sex must be determined by PCR amplification of X chromosome genes with divergent Y chromosome gametologs. Existing simplex PCR methods are confounded by small size differences between amplicons, amplification of unexpected products, and biased amplification of the shorter amplicon. RESULTS: Primers were designed flanking an 84 bp deletion of the X-linked Rbm31x gene relative to its Y-linked gametolog Rbm31y. A single product was amplified from XX samples, with two products amplified from XY samples. Amplicons were resolved by gel electrophoresis for 20 min, with unbiased amplification of both products observed in XY samples. CONCLUSION: This method achieves rapid and unequivocal genetic sex determination of mice in low volume PCR reactions, reducing reagent usage and simultaneously eliminating shortcomings of previous methods.
Rbm31x, Rbm31y, Sex genotyping of mice, Simplex PCR, Animals, Genes, X-Linked, Genes, Y-Linked, Mice, 129 Strain, Mice, Inbred C57BL, Polymerase Chain Reaction, Sex Determination Analysis, Time Factors
External DOI: https://doi.org/10.1186/s13293-017-0154-6
This record's URL: https://www.repository.cam.ac.uk/handle/1810/284759
Attribution 4.0 International
Licence URL: https://creativecommons.org/licenses/by/4.0/
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