Rapid sexing of bovine preimplantation embryos using loop-mediated isothermal amplification

Theriogenology
Hiroki HirayamaAkira Minamihashi

Abstract

Loop-mediated isothermal amplification (LAMP) is a novel DNA amplification method that amplifies a target sequence specifically under isothermal conditions. The product of LAMP is detected by the turbidity of the reaction mixture without electrophoresis. The objective of this study was to develop a rapid sexing method for bovine preimplantation embryos using LAMP. The first experiment was conducted to optimize the DNA extraction method for LAMP-based embryo sexing. The DNA of single blastomeres was extracted using three methods: heat, NaOH, and proteinase K-Tween 20 (PK-TW) treatments. Sexing was performed with two LAMP reactions, male-specific and male-female common reaction, after DNA extraction. The rates of correct determination of sex were 88.9-94.4%, with no difference among methods. The sensitivity and accuracy of LAMP-based embryo sexing were evaluated in the next experiment. The proportion of samples in which the sex was correctly determined was 75-100% for one to five biopsied cells. Lastly, in vivo-derived embryos were examined to verify the usefulness of LAMP-based embryo sexing, and some of these fresh, sexed embryos were transferred into recipient animals. The time needed for sexing was <1 h. The pregnancy rate wa...Continue Reading

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Citations

Jan 13, 2006·BMC Biotechnology·Yasuyoshi MoriTsugunori Notomi
Jan 14, 2005·The Journal of Reproduction and Development·Hiroki HirayamaAkira Minamihashi
Nov 9, 2005·The Journal of Reproduction and Development·Budiyanto AgungTakashi Nagai
Jun 15, 2007·The Journal of Reproduction and Development·Atsushi IdetaYoshito Aoyagi
Nov 3, 2009·The Journal of Reproduction and Development·Hiroki HirayamaAkira Minamihashi
Sep 17, 2010·Applied Biochemistry and Biotechnology·Shijun FuZhiqiang Shen
Oct 12, 2012·World Journal of Microbiology & Biotechnology·Preeti SinghSundeep Singh
Mar 15, 2011·Theriogenology·K M A Zoheir, A A Allam
Mar 23, 2011·Parasitology International·Toshihiro TokiwaNobuo Ohta
Jan 12, 2010·Animal Reproduction Science·Khairy M A Zoheir, Ahmed A Allam
Oct 28, 2008·Theriogenology·H HayakawaY Aoyagi
Dec 17, 2008·Forensic Science International. Genetics·Hiroaki NogamiRei Mukoyama
Nov 21, 2008·Reproduction in Domestic Animals = Zuchthygiene·Z-P ZhangS W J Pu
Dec 2, 2006·International Journal of Cancer. Journal International Du Cancer·Daisuke HoribeHideki Hayashi
Jun 11, 2008·Molecular Reproduction and Development·Florence GuignotPascal Mermillod
Jul 29, 2010·Animal Science Journal = Nihon Chikusan Gakkaihō·Kiyoshi AkiyamaSatoko Matoba
Dec 2, 2011·Reproduction in Domestic Animals = Zuchthygiene·B Wu, L Zan
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Feb 16, 2005·Theriogenology·Hitomi TakahashiKiyoshi Okuda
Mar 26, 2014·The Journal of Reproduction and Development·Koji Kimura, Shuichi Matsuyama
Aug 24, 2013·The Journal of Reproduction and Development·Hiroki HirayamaAkira Minamihashi
Sep 20, 2011·Theriogenology·Navid Ghavi Hossein-Zadeh
Dec 30, 2014·Theriogenology·Trisadee KhamlorNipa Chokesajjawatee
Jun 19, 2016·Bioscience, Biotechnology, and Biochemistry·Junbei XiangLinfeng Xie
Aug 1, 2015·PLoS Neglected Tropical Diseases·Javier GandaseguiAntonio Muro
Dec 6, 2007·Reproduction in Domestic Animals = Zuchthygiene·G A Presicce
Feb 15, 2019·International Journal of Veterinary Science and Medicine·Islam M Saadeldin, Goo Jang
Jun 11, 2019·The Journal of Reproduction and Development·Tomoko ObuchiHitoshi Ushijima

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