LAMP法鉴定家鸡性别的初步研究

Preliminary Investigation of Sex Identification of Gallus by Loop-mediated Isothermal Amplification

为建立一种利用环介导等温扩增(LAMP)技术高效快速鉴定家鸡及早期胚胎性别的反应体系,本研究以W染色体上假基因序列EE 0.6序列为模板,设计LAMP引物,对反应温度、反应时间、Mg2+浓度、d NTP浓度等条件进行优化.结果表明,在25μL反应体系中,Mg2+终浓度为6 mmol/L,d NTP终浓度为1.5 mmol/L,63℃反应60 min时为最佳反应体系.在该反应体系下,LAMP成功地鉴定出家鸡性别的雌雄,准确率达到100%.此方法将成为一种快速有效地鉴定家鸡早期胚胎性别的新方法,且可以在生产实践中广泛推广应用.

A rapid and highly effective detection system was established for gallus and its early-stage embry？os sexing using loop-mediated isothermal amplification （LAMP）. LAMP primers were designed by targeting EE0.6 pseudogene sequence on W chromosome from chicken. The reaction condition of the detection system was optimized by reaction temperature,reaction time and Mg 2＋ and dNTPs concentration. The results showed that the optimal condition was at 63℃for 60 min with 6 mmol/L Mg 2＋and 1.5 mmol/L dNTPs in 25μL reac？tion system. The LAMP method has been successfully applied to detection of adult chicken sexing by optimal system and achieved 100%. This investigation suggested that LAMP will be a rapid method for sex identifi？cation of chicken embryos,and can be extensively applied and popularized in production practice.