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Establishment of New Genetic Markers and Methods for Sex Determination of Mouse and Human Cells using Polymerase Chain Reactions and Crude DNA Samples
Background: The currently available methods for sexing human or mouse cells have weaknesses. Therefore, it is necessary to establish new methods. Methods: We used bioinformatics approach to identify genes that have alleles on both the X and Y chromosomes of mouse and human genomes and have a region...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Bentham Science Publishers
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9875541/ https://www.ncbi.nlm.nih.gov/pubmed/36777874 http://dx.doi.org/10.2174/1389202923666220610121344 |
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author | Zhang, Keyin Yang, Jianglin Qin, Zhenwei Lu, Tianzu Lou, Didong Ran, Qianchuan Huang, Hai Cheng, Shuqiang Zellmer, Lucas Ma, Hong Liao, Dezhong J. |
author_facet | Zhang, Keyin Yang, Jianglin Qin, Zhenwei Lu, Tianzu Lou, Didong Ran, Qianchuan Huang, Hai Cheng, Shuqiang Zellmer, Lucas Ma, Hong Liao, Dezhong J. |
author_sort | Zhang, Keyin |
collection | PubMed |
description | Background: The currently available methods for sexing human or mouse cells have weaknesses. Therefore, it is necessary to establish new methods. Methods: We used bioinformatics approach to identify genes that have alleles on both the X and Y chromosomes of mouse and human genomes and have a region showing a significant difference between the X and Y alleles. We then used polymerase chain reactions (PCR) followed by visualization of the PCR amplicons in agarose gels to establish these genomic regions as genetic sex markers. Results: Our bioinformatics analyses identified eight mouse sex markers and 56 human sex markers that are new, i.e. are previously unreported. Six of the eight mouse markers and 14 of the 56 human markers were verified using PCR and ensuing visualization of the PCR amplicons in agarose gels. Most of the tested and untested sex markers possess significant differences in the molecular weight between the X- and Y-derived PCR amplicons and are thus much better than most, if not all, previously-reported genetic sex markers. We also established several simple and essentially cost-free methods for extraction of crude genomic DNA from cultured cells, blood samples, and tissues that could be used as template for PCR amplification. Conclusion: We have established new sex genetic markers and methods for extracting genomic DNA and for sexing human and mouse cells. Our work may also lend some methodological strategies to the identification of new genetic sex markers for other organismal species. |
format | Online Article Text |
id | pubmed-9875541 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Bentham Science Publishers |
record_format | MEDLINE/PubMed |
spelling | pubmed-98755412023-02-11 Establishment of New Genetic Markers and Methods for Sex Determination of Mouse and Human Cells using Polymerase Chain Reactions and Crude DNA Samples Zhang, Keyin Yang, Jianglin Qin, Zhenwei Lu, Tianzu Lou, Didong Ran, Qianchuan Huang, Hai Cheng, Shuqiang Zellmer, Lucas Ma, Hong Liao, Dezhong J. Curr Genomics Genetics & Genomics Background: The currently available methods for sexing human or mouse cells have weaknesses. Therefore, it is necessary to establish new methods. Methods: We used bioinformatics approach to identify genes that have alleles on both the X and Y chromosomes of mouse and human genomes and have a region showing a significant difference between the X and Y alleles. We then used polymerase chain reactions (PCR) followed by visualization of the PCR amplicons in agarose gels to establish these genomic regions as genetic sex markers. Results: Our bioinformatics analyses identified eight mouse sex markers and 56 human sex markers that are new, i.e. are previously unreported. Six of the eight mouse markers and 14 of the 56 human markers were verified using PCR and ensuing visualization of the PCR amplicons in agarose gels. Most of the tested and untested sex markers possess significant differences in the molecular weight between the X- and Y-derived PCR amplicons and are thus much better than most, if not all, previously-reported genetic sex markers. We also established several simple and essentially cost-free methods for extraction of crude genomic DNA from cultured cells, blood samples, and tissues that could be used as template for PCR amplification. Conclusion: We have established new sex genetic markers and methods for extracting genomic DNA and for sexing human and mouse cells. Our work may also lend some methodological strategies to the identification of new genetic sex markers for other organismal species. Bentham Science Publishers 2022-08-11 2022-08-11 /pmc/articles/PMC9875541/ /pubmed/36777874 http://dx.doi.org/10.2174/1389202923666220610121344 Text en © 2022 Bentham Science Publishers https://creativecommons.org/licenses/by-nc/4.0/ This is an open access article licensed under the terms of the Creative Commons Attribution-Non-Commercial 4.0 International Public License (CC BY-NC 4.0) (https://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited. |
spellingShingle | Genetics & Genomics Zhang, Keyin Yang, Jianglin Qin, Zhenwei Lu, Tianzu Lou, Didong Ran, Qianchuan Huang, Hai Cheng, Shuqiang Zellmer, Lucas Ma, Hong Liao, Dezhong J. Establishment of New Genetic Markers and Methods for Sex Determination of Mouse and Human Cells using Polymerase Chain Reactions and Crude DNA Samples |
title | Establishment of New Genetic Markers and Methods for Sex Determination of Mouse and Human Cells using Polymerase Chain Reactions and Crude DNA Samples |
title_full | Establishment of New Genetic Markers and Methods for Sex Determination of Mouse and Human Cells using Polymerase Chain Reactions and Crude DNA Samples |
title_fullStr | Establishment of New Genetic Markers and Methods for Sex Determination of Mouse and Human Cells using Polymerase Chain Reactions and Crude DNA Samples |
title_full_unstemmed | Establishment of New Genetic Markers and Methods for Sex Determination of Mouse and Human Cells using Polymerase Chain Reactions and Crude DNA Samples |
title_short | Establishment of New Genetic Markers and Methods for Sex Determination of Mouse and Human Cells using Polymerase Chain Reactions and Crude DNA Samples |
title_sort | establishment of new genetic markers and methods for sex determination of mouse and human cells using polymerase chain reactions and crude dna samples |
topic | Genetics & Genomics |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9875541/ https://www.ncbi.nlm.nih.gov/pubmed/36777874 http://dx.doi.org/10.2174/1389202923666220610121344 |
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