Scholarlink

<p>Early sex determination and genotyping of chicken embryos is crucial for ethical and resource-efficient animal research. We developed a reproducible workflow using whole genome amplification combined (WGA) with Kompetitive Allele Specific PCR (KASP) and standard endpoint PCR to perform <i>in ovo</i> sexing and genotyping from embryonic day 4 (ED4) to ED10. Across four study phases involving various chicken lines—including a genetically modified line, Araucana crossbreeds, and commercial layers (total <i>n</i> = 819)—we evaluated methodological feasibility, accuracy, and hatchability. Both PCR methods demonstrated high success rates in yielding results (70–100%) and high identification accuracy (92–100%) throughout all developmental stages tested. While the overall efficiency improved with embryonic age, ED7 was identified as the optimal time point to balance reliable sampling with high hatchability. We conclude that this laboratory-scale, PCR-based protocol provides an accessible and precise method for research laboratories to identify sex and genotype simultaneously. This approach directly supports the 3R principles by preventing the hatching of surplus animals before the assumed onset of nociception (ED13).</p>

错误：搜索内容不能为空，请输入英文关键词
错误：关键词超出字数限制，请精简
高级检索

In ovo sexing and genotyping using PCR techniques: a contribution to the 3R principles in chicken breeding

  • 期刊论文

  • 出版模式：

    开放获取

  • 发表日期：

    2026年2月21日

查看全文
  • C. Dierks,
  • A. Förster,
  • D. Meunier,
  • R. Preisinger,
  • C. Klein,
  • S. Weigend,
  • S. Altgilbers

摘要

Early sex determination and genotyping of chicken embryos is crucial for ethical and resource-efficient animal research. We developed a reproducible workflow using whole genome amplification combined (WGA) with Kompetitive Allele Specific PCR (KASP) and standard endpoint PCR to perform in ovo sexing and genotyping from embryonic day 4 (ED4) to ED10. Across four study phases involving various chicken lines—including a genetically modified line, Araucana crossbreeds, and commercial layers (total n = 819)—we evaluated methodological feasibility, accuracy, and hatchability. Both PCR methods demonstrated high success rates in yielding results (70–100%) and high identification accuracy (92–100%) throughout all developmental stages tested. While the overall efficiency improved with embryonic age, ED7 was identified as the optimal time point to balance reliable sampling with high hatchability. We conclude that this laboratory-scale, PCR-based protocol provides an accessible and precise method for research laboratories to identify sex and genotype simultaneously. This approach directly supports the 3R principles by preventing the hatching of surplus animals before the assumed onset of nociception (ED13).