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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...

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Autores principales: FUJIMOTO, Masaki, KITAMURA, Hiroshi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: The Japanese Society of Veterinary Science 2022
Materias:
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.
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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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