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Genetic sex determination of mice by simplex PCR
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 de...
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2017
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5645908/ https://www.ncbi.nlm.nih.gov/pubmed/29041956 http://dx.doi.org/10.1186/s13293-017-0154-6 |
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author | Tunster, Simon James |
author_facet | Tunster, Simon James |
author_sort | Tunster, Simon James |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-5645908 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-56459082017-10-26 Genetic sex determination of mice by simplex PCR Tunster, Simon James Biol Sex Differ Letter to the Editor 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. BioMed Central 2017-10-17 /pmc/articles/PMC5645908/ /pubmed/29041956 http://dx.doi.org/10.1186/s13293-017-0154-6 Text en © The Author(s). 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Letter to the Editor Tunster, Simon James Genetic sex determination of mice by simplex PCR |
title | Genetic sex determination of mice by simplex PCR |
title_full | Genetic sex determination of mice by simplex PCR |
title_fullStr | Genetic sex determination of mice by simplex PCR |
title_full_unstemmed | Genetic sex determination of mice by simplex PCR |
title_short | Genetic sex determination of mice by simplex PCR |
title_sort | genetic sex determination of mice by simplex pcr |
topic | Letter to the Editor |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5645908/ https://www.ncbi.nlm.nih.gov/pubmed/29041956 http://dx.doi.org/10.1186/s13293-017-0154-6 |
work_keys_str_mv | AT tunstersimonjames geneticsexdeterminationofmicebysimplexpcr |