Sexing and multiple genotype analysis from a single cell of bovine embryo

Theriogenology
P ChrenekJ P Renard

Abstract

We described a procedure for multiple genotype analysis (determination of sex and of three genetic markers) from a single cell derived from bovine preimplantation embryo. It consists of primer extension preamplification-polymerase chain reaction (PEP-PCR) and subsequent single assay or multiplex PCR. A single blastomere that was isolated by microaspiration from bovine embryos at the 16- to 32-cell stage then was lysed and was subjected to the PEP-PCR. When testing 75 embryos, efficiency of genotyping by standard PCR for kappa-casein, growth hormone (GH) and prolactin (PRL) polymorphic alleles was 91, 88 and 89%, respectively. Sexing efficiency in the multiplex PCR was 91%, based on the amplification of Y-specific locus using kappa-casein internal standard. The microaspiration of a single blastomere was shown not to be invasive for the embryos. It did not alter their development potential in vitro (P > 0.05), as was seen by obtaining a similar percentage of embryos developing further into the blastocyst stage in the group subjected to biopsy (44/75, 59%) and in the control group of embryos (30/50, 60%).

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Citations

Mar 24, 2004·Theriogenology·Keiichiro Tominaga, Yukako Hamada
Jul 15, 2004·Theriogenology·Hiroki HirayamaAkira Minamihashi
Apr 15, 2008·Zygote : the Biology of Gametes and Early Embryos·Francesca CianiRoberto Lorizio
Jan 28, 2003·Nature Biotechnology·Brigid BrophyGötz Laible
Jul 23, 2004·Cloning and Stem Cells·D Bousquet, P Blondin
Nov 3, 2009·The Journal of Reproduction and Development·Hiroki HirayamaAkira Minamihashi
Jan 14, 2005·The Journal of Reproduction and Development·Hiroki HirayamaAkira Minamihashi
Dec 19, 2015·Systems Biology in Reproductive Medicine·Tahereh HaghpanahHadi Azimi
Mar 15, 2011·Theriogenology·K M A Zoheir, A A Allam
Jan 12, 2010·Animal Reproduction Science·Khairy M A Zoheir, Ahmed A Allam
Jul 28, 2004·Molecular Reproduction and Development·L MaraP Cappai
Jan 28, 2003·Molecular Reproduction and Development·E M JoudreyW A King
Sep 12, 2007·Reproduction in Domestic Animals = Zuchthygiene·H M Kubisch, K M Johnson
Oct 29, 2005·Journal of Experimental Zoology. Part A, Comparative Experimental Biology·Valerie J Grant, R J Irwin
Apr 19, 2007·Molecular Reproduction and Development·Jaana PeippoJohanna Vilkki
Jun 11, 2008·Molecular Reproduction and Development·Florence GuignotPascal Mermillod
Dec 3, 2010·Reproduction in Domestic Animals = Zuchthygiene·F GuignotG Baril
Sep 29, 2004·Theriogenology·Yan-Ming HorngMu-Chiou Huang
Oct 24, 2012·Journal of Biomedicine & Biotechnology·M CenariuI Groza
May 9, 2006·The Journal of Veterinary Medical Science·Soichi KageyamaSadao Onoe
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
Dec 30, 2014·Theriogenology·Trisadee KhamlorNipa Chokesajjawatee
Sep 15, 2020·Frontiers in Veterinary Science·Yanshe XieLinjun Hong

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