Journal of Embryo Transfer (한국수정란이식학회지)

The Korean Society of Embryo Transfer (한국수정란이식학회)
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The Improvement of Sexing PCR Conditions and Survival Rate of Blastomere Separation Method in the Bovine Embryo

소 수정란의 할구 분리방법에 따른 생존율 및 성판별 PCR의 개선

  • Kim, Sang-Hwan (Institute of Genetic Engineering, Hankyong National University) ;
  • Kim, Kyong-Lae (Institute of Genetic Engineering, Hankyong National University) ;
  • Lee, Ho-Jun (Institute of Genetic Engineering, Hankyong National University) ;
  • Jung, Kyoung-Sub ;
  • Baek, Jun-Seok ;
  • Jung, Duk-Won (Major in Animal Fusion Biotechnology, Graduate School of Bio & Information Technology, Hankyong National University) ;
  • Kim, Dae-Eun (Department of Bio-Medical Science Laboratory, Kyungbok University) ;
  • Lee, Deuk-Hwan (Department of Animal Life Science, Hankyong National University) ;
  • Yoon, Jong-Taek (Institute of Genetic Engineering, Hankyong National University)
  • 김상환 (한경대학교 유전공학연구소) ;
  • 김경래 (한경대학교 유전공학연구소) ;
  • 이호준 (한경대학교 유전공학연구소) ;
  • ;
  • ;
  • 정덕원 (한경대학교 미래융합기술대학원 동물바이오융합전공) ;
  • 김대은 (경복대학교 임상병리학과) ;
  • 이득환 (한경대학교 동물생명환경과학부 동물생명공학전공) ;
  • 윤종택 (한경대학교 유전공학연구소)
  • Received : 2013.08.23
  • Accepted : 2013.09.12
  • Published : 2013.09.30
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Abstract

The present study was conducted to compare on embryo survival rates by blastomere isolation methods, and establish the optimal PCR procedure for perform the sexing of bovine blastocysts produced by IVF. IVF embryos used in the study was used the Bisected or Sliced methods for blastomere isolation, and the survival rates of blastocyst with rapid way of sexing PCR was assessed. In the present study for survival rates in blastocyst was the total cleavage rate was 75% and a blastocyst development among cleaved embryos was 40%. Survival rate of embryos treated with intact, bisected or sliced method was 100, 63.3 or 81.3%, respectively. Therefore, survival rate of embryos treated with sliced method was higher compared to that of embryos treated with bisected method. The sexing rate of female or male was not significantly different between S4BFBR primer and BSY + BSP primer (1.75 : 1 vs. 1.43 : 1), respectively. Because of the PCR amplification using the S4BFBR primer was simpler method than multiplex PCR amplification method. Furthermore, the accuracy of sexing rate and reduction of PCR work time between 2-step and 3-step of PCR methods was 98.0% / 1.5 hr and 97.0% / 3.5 hr, respectively. Based on these results, it can be suggested that the sliced and PCR methods we developed was very effective method to reduce time consuming and procedure of PCR amplification for sexing with the increase of survival rate on the blastocyst.

Keywords

References

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