一种简单高效稳定的高质量第X期鸡胚盘DNA提取方法的建立

Establishment of a Simple, Efficient and Stable Method for DNA Extraction from Stage X Chicken Blastoderm

为了建立快速高效稳定的高质量鸡胚盘DNA提取方法,试验通过收集第X期鸡胚胎,应用酚氯仿抽提法、微量组织DNA提取试剂盒法和优化的硅胶膜吸附法分别提取鸡胚DNA,所提DNA经0.8%的琼脂糖凝胶电泳初步检测,并经PCR进一步分析检验,对不同方法提取DNA的稳定性和质量进行评价。结果表明:微量组织DNA提取试剂盒法提取的基因组DNA比用酚氯仿抽提法提取DNA的方法具有更高的稳定性和更好的质量,而微量组织DNA提取试剂盒法与本试验建立的优化的硅胶膜吸附法提取的基因组DNA稳定性和质量相近,但本试验建立优化的硅胶膜吸附法,无需蛋白酶K消化胚盘组织,因而优化了步骤降低了成本。试验建立了一个简单高效稳定的高质量低成本的第X期鸡胚盘DNA提取方法,为后续的相关研究建立了一个有效的鸡受精蛋胚盘DNA提取方法。

In order to establish a rapid, efficient, stable and high quality chicken blastoderm DNA extraction method, the X stage chicken blastoderm were collected and extracted by phenol chloroform extraction method,Kit Method of DNA extraction from micro tissue and optimized silicone membrane adsorption method respectively. The stability and quality of the extracted DNA were preliminary evaluated by agarose gel electrophoresis of 0.8% and further analyzed by PCR. The results showed that the genomic DNA extracted by Kit Method of DNA extraction from micro tissue was more stable and better quality than that extracted by phenol chloroform extraction method, while the stability and quality of genomic DNA extracted by Kit Method of DNA extraction from micro tissue was similar to that of the optimized silicone membrane adsorption method. However, the optimized silicone membrane adsorption method was established, which did not need protease K to digest the blastoderm tissue, so the steps were optimized and the cost was reduced. The results showed that a simple, efficient and stable DNA extraction method with high-quality and low-cost for X stage chicken blastoderm were established and it can be used in subsequent related studies.