title: Embryo sexing and sex chromosomal chimerism analysis by loop-mediated isothermal amplification in cattle and water buffaloes.
abstract: In domestic animals of the family Bovidae, sex preselection of offspring has been demanded for convenience of milk/beef production and animal breeding. Development of the nonsurgical embryo transfer technique and sexing methods of preimplantation embryos made it possible. Sexing based on detection of Y chromosome-specific DNA sequences is considered the most reliable method to date. PCR enables amplification of a target sequence from a small number of blastomeres. However, it requires technical skill and is time consuming. Furthermore, PCR has the risk of false positives because of DNA contamination during handling of the PCR products in duplicate PCR procedures and/or electrophoresis. Therefore, for embryo sexing to become widely used in the cattle embryo transfer industry, a simple, rapid and precise sexing method needs to be developed. Loop-mediated isothermal amplification (LAMP) is a novel DNA amplification method, and the reaction is carried out under isothermal conditions (range, 60 to 65 C) using DNA polymerase with strand displacement activity. When the target DNA is amplified by LAMP, a white precipitate derived from magnesium pyrophosphate (a by-product of the LAMP reaction) is observed. It is noteworthy that LAMP does not need special reagents or electrophoresis to detect the amplified DNA. This review describes the development and application of an embryo sexing method using LAMP in cattle and water buffaloes.
authors: Animal Biotechnology Group, Animal Research Center, Hokkaido Research Organization, Hokkaido 081-0038, Japan.
authors: Hiroki
authors: H
authors: Hirayama
authors: Soichi
authors: S
authors: Kageyama
authors: Satoru
authors: S
authors: Moriyasu
authors: Ken
authors: K
authors: Sawai
authors: Akira
authors: A
authors: Minamihashi
mesh_terms: D000818:Animals; D002020:Buffaloes / Q000235:genetics / Q000502:physiology*; D002417:Cattle / Q000235:genetics / Q000502:physiology*; D005260:Female; D021141:Nucleic Acid Amplification Techniques / Q000379:methods / Q000662:veterinary*; D012732:Sex Determination Analysis / Q000379:methods / Q000662:veterinary*; D014998:Y Chromosome
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