Application of the colorimetric loop-mediated isothermal amplification (LAMP)
technique for genotyping Cre-driver mice |
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| Authors: | Masaki FUJIMOTO Hiroshi KITAMURA |
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| Institution: | 1)Laboratory of Radiation Biology, Faculty of Veterinary Medicine, Hokkaido University, Hokkaido, Japan;2)Laboratory of Veterinary Physiology, Department of Veterinary Medicine, School of Veterinary Medicine, Rakuno Gakuen University, Hokkaido, Japan |
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| Abstract: | 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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| Keywords: | conditional knockout Cre genotyping loop-mediated isothermal amplification |
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