Two-Color Allele-Specific Polymerase Chain Reaction (PCR-SSP) Assay of the Leptin Receptor Gene (Lepr^<db>) for Genotyping Mouse Diabetes Mutation
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概要
- 論文の詳細を見る
An allele specific polymerase chain reaction (PCR-SSP) assay for genotyping the mouse leptin receptor (Leprdb) mutation and its wild type (Lepr+) gene was developed using two different fluorescent dye-labeled primers. First, we determined the Leprdb and Lepr+ allele by PCR-SSP assay with usual dye-unlabeled primers. However this method requires two separate PCR reactions because the amplified products specific for each allele are almost the same size. We further developed a simple and reliable two-color PCR-SSP method that uses a color complementation strategy to distinguish the Lepr db and Lepr+ alleles. Leprdb/Lepr db, Leprdb/Lepr+ and Lepr+/Lepr+ of mice (5 each ) were clearly genotyped by the two-color PCR-SSP. We also performed PCR-direct sequencing for the same samples and confirmed the accuracy of this method. This method makes it possible to reduce the number of PCR reactions because both alleles are amplified in the same reaction mixture.
- 社団法人 日本実験動物学会の論文
- 2001-10-01
著者
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SUEMIZU Hiroshi
Central Institute for Experimental Animals
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Ohishi Yumiko
東京大学 医学系研究科循環器内科
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Tamaki Norikazu
Central Institute For Experimental Animals
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OHNISHI Yasuyuki
Central Institute for Experimental Animals
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TAMAOKI Norikazu
Central Institute for Experimental Animals
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MARUYAMA Chika
Central Institute for Experimental Animals
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Ohnishi Yasuyuki
Biomedical Research Department Central Institute For Experimental Animals
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Tamaoki N
Central Institute For Experimental Animals
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Tamaoki Norikazu
Central Inst. For Experimental Animals
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KITO MARUYAMA
Central Institute for Experimental Animals
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Ohnishi Yasuyuki
Central Inst. For Experimental Animals
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