Development of an Efficient Genotyping Method to Detect Obese Mutation in the Mouse Leptin Gene for Use in SPF Barrier Facilities
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- AYABE Hiroaki
- Production Department, Plasma Team, CHARLES RIVER LABORATORIES JAPAN, Inc., 795 Shimofurusawa, Atsugi-shi, Kanagawa 243–0214, Japan
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- IKEDA Shogo
- Department of Biochemistry, Faculty of Science, Okayama University of Science, 1–1 Ridaicho, Kita-ku, Okayama-shi, Okayama 700–0005, Japan
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- MARUYAMA Shigeri
- Monitoring Center, CHARLES RIVER LABORATORIES JAPAN, Inc., 4049–3 Nakatsu, Aikawa-machi, Aikou-gun, Kanagawa 243–0303, Japan
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- SHIOYAMA Seiji
- Tsukuba Breeding Center, CHARLES RIVER LABORATORIES JAPAN, Inc., 955 Kamibayashi, Ishioka-shi, Ibaraki 315–0138, Japan
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- KIKUCHI Mayumi
- Monitoring Center, CHARLES RIVER LABORATORIES JAPAN, Inc., 4049–3 Nakatsu, Aikawa-machi, Aikou-gun, Kanagawa 243–0303, Japan
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- KAWAGUCHI Ayako
- Monitoring Center, CHARLES RIVER LABORATORIES JAPAN, Inc., 4049–3 Nakatsu, Aikawa-machi, Aikou-gun, Kanagawa 243–0303, Japan
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- YAMADA Tetsuo
- Hino Breeding Center, CHARLES RIVER LABORATORIES JAPAN, Inc., 735 Shimokomaduki, Hino-cho, Gamou-gun, Shiga 529–1633, Japan
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- IKEDA Takuya
- Production Department, CHARLES RIVER LABORATORIES JAPAN, Inc., 3–17–6 Shin-Yokohama, Kohoku-ku, Yokohama-shi, Kanagawa 222–0033, Japan
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Abstract
We have developed a rapid and efficient genotyping method for detection of the mouse leptin obese mutation (Lepob) using tetra-primer amplification refractory mutation system-polymerase chain reaction (tetra-primer ARMS-PCR). In this method, whole blood collected onto gamma-ray sterilized Flinders Technology Associates (FTA) filter paper is used as PCR template without a DNA purification step. Three genotypes (Lepob/Lepob, Lepob/+ and +/+) differentiated by single-tube PCR and electrophoresis were perfectly consistent with those determined by PCR-restriction fragment length polymorphism (PCR-RFLP). This method can save material costs and operation time, because it does not require restriction enzyme digestion and could be set up in most specific pathogen-free (SPF) barrier facilities.
Journal
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- Journal of Veterinary Medical Science
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Journal of Veterinary Medical Science 75 (5), 633-638, 2013
JAPANESE SOCIETY OF VETERINARY SCIENCE
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Details 詳細情報について
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- CRID
- 1390282681404536960
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- NII Article ID
- 130002502167
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- NII Book ID
- AA10796138
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- COI
- 1:STN:280:DC%2BC3s3gtFyqtg%3D%3D
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- ISSN
- 13477439
- 09167250
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- NDL BIB ID
- 024676371
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- PubMed
- 23229540
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- Text Lang
- en
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- Data Source
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- JaLC
- NDL
- Crossref
- PubMed
- CiNii Articles
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- Abstract License Flag
- Disallowed