| 适配子筛选中随机寡核苷酸文库的构建和PCR条件的优化简 |
| |
| 引用本文： | 田瑞云,卢士英,李琳,吴宗澄,任洪林,周玉,李岩松,胡盼,柳增善,王光明.适配子筛选中随机寡核苷酸文库的构建和PCR条件的优化简[J].中国畜牧兽医,2014,41(1):61-65. |
| |
| 作者姓名： | 田瑞云 卢士英 李琳 吴宗澄 任洪林 周玉 李岩松 胡盼 柳增善 王光明 |
| |
| 作者单位： | (人兽共患病研究教育部重点实验室，吉林大学人兽共患病研究所，吉林大学动物医学学院，吉林长春 130062) |
| |
| 基金项目： | 国家自然科学基金（31071539）。 |
| |
| 摘 要： | 本试验旨在构建用于核酸适配子筛选的随机寡核苷酸文库并对筛选过程中的PCR条件进行优化。利用Primer Premier 5.0构建了随机寡核苷酸文库并据文库两端固定序列设计引物，对对称PCR和间接不对称PCR中的上、下游引物的比例、模板量、退火温度、循环数等条件进行优化并验证其重复性，得到了对称PCR和间接不对称PCR的最佳反应条件，获得较理想的dsDNA和ssDNA并具有良好的重复性。本试验成功构建了寡核苷酸文库，优化了PCR条件，为适配子筛选和后续试验奠定了一定的基础。
|
| 关 键 词： | 适配子 寡核苷酸文库 PCR 优化 |
Construction of Oligonucleotide Library in Selection of Aptamer and Optimization of PCR Conditions |
| |
| TIAN Rui-yun,LU Shi-ying,LI Lin,WU Zong-cheng,REN Hong-lin,ZHOU Yu,LI Yan-song,HU Pan,LIU Zeng-shan,WANG Guang-ming.Construction of Oligonucleotide Library in Selection of Aptamer and Optimization of PCR Conditions[J].China Animal Husbandry & Veterinary Medicine,2014,41(1):61-65. |
| |
| Authors: | TIAN Rui-yun LU Shi-ying LI Lin WU Zong-cheng REN Hong-lin ZHOU Yu LI Yan-song HU Pan LIU Zeng-shan WANG Guang-ming |
| |
| Affiliation: | （Key Laboratory of Zoonosis Research, Ministry of Education， Institute of Zoonosis， College of Veterinary Medicine, Jilin University，Changchun 130062, China） |
| |
| Abstract: | The purpose of this experiment was to construct the oligonucleotide library used in the selection of aptamer and optimize the PCR conditions in the selection process. We constructed the oligonucleotide library and designed primers according to the immobilized sequence of amphi-oligonucleotide using Primer Premier 5.0. The proportion of upstream and downstream primers, quantity of template, anneal temperature and cycle numbers of the symmetric and asymmetric PCR were optimized and their repeatabilities were verified. After optimizing, we set up the best symmetric and asymmetric PCR conditions achieved the desirable dsDNA and ssDNA. The optimized PCR showed high reproducibility. We constructed the oligonucleotide library and optimized PCR condition successfully. This study provided a reference for selection of aptamer and laid a foundation for further study. |
| |
| Keywords: | aptamer oligonucleotide library PCR optimization |
| 本文献已被 CNKI 等数据库收录！ |
| 点击此处可从《中国畜牧兽医》浏览原始摘要信息 |
|
点击此处可从《中国畜牧兽医》下载全文 |
|
