延边黄牛体细胞克隆囊胚中总基因组RNA的提取

RNA Extraction from Blastocyst of Somatic Cell Cloning in Yanbian Yellow Cattle

以延边黄牛体细胞克隆囊胚为试验材料,采用试剂盒GMS12106的使用方法提取试材的总基因组RNA,在琼脂糖凝胶电泳检测后,得到最适的囊胚数为5个。通过试验发现,生殖细胞的特殊生理结构限制了RNA提取的效果。为提取完整、纯度高的RNA,采用链蛋白酶对囊胚的透明带进行了消化处理,最终得出链蛋白酶处理的最适时间为10min,提取的RNA质量好,可为后续试验奠定基础。

In this study, RNA extraction kit （GMS12106） was used in RNA extraction from blastocyst of somatic cell cloning in Yanbian yellow cattle. The detecting result by agarose gel electrophoresis proved that five blastocysts were obtained. The results of the experiments showed that the effectiveness of RNA extraction was limited because of the special physiological structure of the reproductive cells. The zona pellucida of the blastocyst was digested with protase in our experiments to extract complete, high-purity RNA. Finally, the results showed that the most appropriate time for the treatment of protase was 10 rain, and the quality of the extracted RNA was better. It laid the foundation for further test.