一种优化的提高质粒产量的提取方法  

An optimized method to improve plasmid DNA extraction efficiency

作　　者：王宗宇 张晓瑛 张小玲 杨永广[1,2] 代相鹏[1,2] WANG Zongyu;ZHANG Xiaoying;ZHANG Xiaoling;YANG Yongguang;DAI Xiangpeng(National-Local Joint Engineering Laboratory of Animal Models for Human Diseases,First Hospital,Jilin University,Changchun 130021,China;Key Laboratory of Organ Regeneration and Transplantation of Ministry of Education,First Hospital,Jilin University,Changchun 130021,China)

机构地区：[1]吉林大学第一医院人类疾病动物模型国家地方联合工程实验室,吉林长春130021 [2]吉林大学第一人民医院器官再造与移植教育部重点实验室,吉林长春130021

出　　处：《畜牧与兽医》2021年第11期42-47,共6页Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金资助项目(81941008);国家自然科学基金面上项目(81972558);吉林省自然科学基金(学科布局项目)(20200201367JC)。

摘　　要：研究旨在优化质粒提取方法,提高质粒产量。对商品化质粒提取试剂盒的提取条件进行了优化,增加了提前小管摇菌和异丙醇-20℃过夜沉淀两个步骤。利用紫外分光光度计、琼脂糖凝胶电泳、双酶切和免疫印迹等方法,对获得质粒的生化特性和生物学功能进行了测定。结果表明,优化的质粒提取方法可提高30%的质粒产量,质粒的生化指标和生物学功能试验结果显示优化方法获得的质粒质量没有下降。The aim of this study was to increase extraction yield of plasmids DNA by optimizing the extraction method. The protocol of a commercial plasmid extraction kit was optimized here by adding two important steps: pre-incubation of competent cells in small tubes and overnight precipitation of isopropanol at-20 ℃. Then, the biochemical characteristics and biological functions of the extracted plasmids DNA were measured by UV spectrophotometer, agarose gel electrophoresis, double enzyme digestion and western blot. The results showed that the extraction yield of plasmids DNA using the optimized method was increased by 30 percent. And the purity of the plasmids DNA, their status, response to double enzymes digestion and expression efficacy in HEK293 cells and PFF were not negatively affected which indicated that the optimized method was successfully established in this study.

关 键 词：质粒提取 方法优化 异丙醇 

分 类 号：R392.3[医药卫生—免疫学]