Tavares K C S, Carneiro I S, Rios D B, Feltrin C, Ribeiro A K C, Gaudêncio-Neto S, Martins L T, Aguiar L H, Lazzarotto C R, Calderón C E M, Lopes F E M, Teixeira L P R, Bertolini M, Bertolini L R
Laboratório de Biologia Molecular e do Desenvolvimento, Núcleo de Biologia Experimental, Universidade de Fortaleza, Fortaleza, CE, Brasil.
Laboratório de Embriologia e Biotecnologia Reprodutiva, Escola de Medicina Veterinária, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brasil.
Genet Mol Res. 2016 Mar 22;15(1):gmr7476. doi: 10.4238/gmr.15017476.
Embryo sexing is a powerful tool for livestock producers because it allows them to manage their breeding stocks more effectively. However, the cost of supplies and reagents, and the need for trained professionals to biopsy embryos by micromanipulation restrict the worldwide use of the technology to a limited number of specialized groups. The aim of this study was to couple a fast and inexpensive DNA extraction protocol with a practical biopsy approach to create a simple, quick, effective, and dependable embryo sexing procedure. From a total of 1847 sheep and cattle whole embryos or embryo biopsies, the sexing efficiency was 100% for embryo biopsies, 98% for sheep embryos, and 90.2% for cattle embryos. We used a primer pair that was common to both species and only 10% of the total extracted DNA. The whole protocol takes only 2 h to perform, which suggests that the proposed procedure can be readily applied to field conditions. Moreover, in addition to embryo sexing, the procedure can be used for further analyses, such as genotyping and molecular diagnosis in preimplantation embryos.
胚胎性别鉴定对畜牧生产者来说是一项强大的工具,因为它能让他们更有效地管理繁殖种群。然而,耗材和试剂的成本,以及需要训练有素的专业人员通过显微操作对胚胎进行活检,限制了这项技术在全球范围内仅被少数专业群体使用。本研究的目的是将一种快速且廉价的DNA提取方案与一种实用的活检方法相结合,以创建一种简单、快速、有效且可靠的胚胎性别鉴定程序。在总共1847个绵羊和牛的完整胚胎或胚胎活检样本中,胚胎活检的性别鉴定效率为100%,绵羊胚胎为98%,牛胚胎为90.2%。我们使用了一对两种物种通用的引物,且仅使用了提取的总DNA的10%。整个方案执行仅需2小时,这表明所提出的程序可以很容易地应用于实际情况。此外,除了胚胎性别鉴定外,该程序还可用于进一步的分析,如植入前胚胎的基因分型和分子诊断。