家鸡囊胚细胞微量DNA提取方法的探讨

Exploration and discussion of Mini-extraction DNA for chicken blastulan cell

为了提取微量的家鸡囊胚细胞DNA应用于种蛋孵化前的PCR性别鉴定,本实验应用Chelex-100法、常规试剂提取法、DNA试剂盒提取法进行微量囊胚细胞的DNA提取,并应用W染色体性别鉴定引物进行PCR鉴定。结果表明,3种方法均能提取适合于PCR的模板,其中以Chelex-100法提取的效果最好,最小细胞数为10个,其次为常规试剂提取法和DNA试剂盒提取法,最小细胞数分别为50个和1200个。本研究对微量家鸡囊胚细胞DNA提取方法进行了大量的摸索和改进,为家鸡种蛋孵化前性别鉴定技术研究奠定了一定的基础。

For extract mini-DNA of chicken blastulan cell to identify sex by PCR, this article extract mini-DNA by chelex-100 extraction, normal reagent extraction and DNA isolation Kit, and apply specific PCR technology to identify extraction result by prime of W chromosome. The results showed that mini-DNA was extracted by different method is adapt to PCR as template, and method of Chelex-100 has best result for PCR with ten cell as template, the next were normal reagent-extraction and DNA isolation Kit, respective to 50 cell and 1200 cell. The research Explored and developed the technology of mini-DNA extraction by different method, it is a base for sex identification in poultry egg without hatch.