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Application of the colorimetric loop-mediated isothermal amplification (LAMP) technique for genotyping Cre-driver mice
The Cre-loxP system is widely used to investigate the cell-type specific roles of genes of interest. Cre-driver mice are required for cell-type specific knockout during the Cre-loxP reaction. To maintain Cre-driver mouse strains, Polymerase chain reaction (PCR)-oriented genotyping targeting the Cre...
Autores principales: | , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
The Japanese Society of Veterinary Science
2022
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9096032/ https://www.ncbi.nlm.nih.gov/pubmed/35228407 http://dx.doi.org/10.1292/jvms.21-0658 |
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author | FUJIMOTO, Masaki KITAMURA, Hiroshi |
author_facet | FUJIMOTO, Masaki KITAMURA, Hiroshi |
author_sort | FUJIMOTO, Masaki |
collection | PubMed |
description | The Cre-loxP system is widely used to investigate the cell-type specific roles of genes of interest. Cre-driver mice are required for cell-type specific knockout during the Cre-loxP reaction. To maintain Cre-driver mouse strains, Polymerase chain reaction (PCR)-oriented genotyping targeting the Cre gene cassette is usually conducted. In this study, we instead applied a colorimetric loop-mediated isothermal amplification (LAMP) method for Cre-genotyping. Among four sets of primers designed by the in silico program, one set effectively amplified the Cre cassette of three Cre-driver strains, but not of C57BL/6 mice. This LAMP-oriented method reduces assay time by less than half compared to the PCR-based method, and can be carried out using a conventional isothermal incubator. Applying this LAMP method may accelerate genotyping of Cre-driver mice. |
format | Online Article Text |
id | pubmed-9096032 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | The Japanese Society of Veterinary Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-90960322022-05-18 Application of the colorimetric loop-mediated isothermal amplification (LAMP) technique for genotyping Cre-driver mice FUJIMOTO, Masaki KITAMURA, Hiroshi J Vet Med Sci Laboratory Animal Science The Cre-loxP system is widely used to investigate the cell-type specific roles of genes of interest. Cre-driver mice are required for cell-type specific knockout during the Cre-loxP reaction. To maintain Cre-driver mouse strains, Polymerase chain reaction (PCR)-oriented genotyping targeting the Cre gene cassette is usually conducted. In this study, we instead applied a colorimetric loop-mediated isothermal amplification (LAMP) method for Cre-genotyping. Among four sets of primers designed by the in silico program, one set effectively amplified the Cre cassette of three Cre-driver strains, but not of C57BL/6 mice. This LAMP-oriented method reduces assay time by less than half compared to the PCR-based method, and can be carried out using a conventional isothermal incubator. Applying this LAMP method may accelerate genotyping of Cre-driver mice. The Japanese Society of Veterinary Science 2022-02-27 2022-04 /pmc/articles/PMC9096032/ /pubmed/35228407 http://dx.doi.org/10.1292/jvms.21-0658 Text en ©2022 The Japanese Society of Veterinary Science https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives (by-nc-nd) License. (CC-BY-NC-ND 4.0: https://creativecommons.org/licenses/by-nc-nd/4.0/) |
spellingShingle | Laboratory Animal Science FUJIMOTO, Masaki KITAMURA, Hiroshi Application of the colorimetric loop-mediated isothermal amplification (LAMP) technique for genotyping Cre-driver mice |
title | Application of the colorimetric loop-mediated isothermal amplification (LAMP)
technique for genotyping Cre-driver mice |
title_full | Application of the colorimetric loop-mediated isothermal amplification (LAMP)
technique for genotyping Cre-driver mice |
title_fullStr | Application of the colorimetric loop-mediated isothermal amplification (LAMP)
technique for genotyping Cre-driver mice |
title_full_unstemmed | Application of the colorimetric loop-mediated isothermal amplification (LAMP)
technique for genotyping Cre-driver mice |
title_short | Application of the colorimetric loop-mediated isothermal amplification (LAMP)
technique for genotyping Cre-driver mice |
title_sort | application of the colorimetric loop-mediated isothermal amplification (lamp)
technique for genotyping cre-driver mice |
topic | Laboratory Animal Science |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9096032/ https://www.ncbi.nlm.nih.gov/pubmed/35228407 http://dx.doi.org/10.1292/jvms.21-0658 |
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