| 信号芋螺基因组DNA提取方法的优化 |
| |
| 引用本文： | 陈婉姨,李新佳,罗素兰,长孙东亭.信号芋螺基因组DNA提取方法的优化[J].热带生物学报,2020,11(2):238. |
| |
| 作者姓名： | 陈婉姨 李新佳 罗素兰 长孙东亭 |
| |
| 作者单位： | 海南大学 热带生物资源教育部重点实验室/生命科学与药学院/海口市海洋药物重点实验室，海口 570228 |
| |
| 基金项目： | 国家自然科学基金项目（41966003，81872794）；高等学校学科创新引智计划项目（D20010） |
| |
| 摘 要： | 高质量的基因组DNA的获取是芋螺毒素基因克隆及基因组文库构建的关键。笔者采用4种方法提取信号芋螺（Conus litteratus)的不同组织器官的DNA，并综合比较了各组DNA的浓度和纯度、DNA片段的大小和完整性。结果表明，由改良的DNA快速提取试剂盒法提取的DNA纯度和浓度较好；4种组织器官中，腹足纯度较高，是更合适的提取组织；肝胰脏和毒管获取的DNA产率最高，但片段完整性差，污染严重；PCR反应获得了跟预期大小一致的特异扩增产物。因此，信号芋螺基因组DNA的提取最佳方法为改良的DNA快速提取试剂盒法，提取部位为腹足组织，可以获得高质量的基因组DNA，且符合后续实验要求。
|
| 关 键 词： | 信号芋螺 基因组DNA 提取方法 优化 |
| 收稿时间： | 2020-02-26 |
Optimization of Genomic DNA Extraction for Conus litteratus |
| |
| Affiliation: | Ministry of Education Key Laboratory of Tropical Biological Resources/Haikou Key Laboratory for Marine Drugs/School of Life Science and Pharmacy, Hainan University, Haikou, Hainan 570228, Hainan, China |
| |
| Abstract: | The acquisition of high-quality genomic DNA is key to cloning of conotoxin genes and gene library construction. Four kits, Magnetic Bead Method Kit, Improved DNA Rapid Extraction Kit, FastPure DNA Isolation Kit, and TIANamp DNA Isolation Kit, were adopted to extract DNA from different tissues and organs of Conus litteratus, and the concentration, purity, size and integrity of the genomic DNA extracted from different tissues and organs by each kit were comprehensively compared. The results showed that the purity and concentration of the genomic DNA extracted with the Improved DNA Rapid Extraction Kit was better. Of the four different organs of C. litterantus, muscular foot had a higher purity in the genomic DNA extracted from its tissues and was the best organ used for extraction of the genomic DNA. The hepatopancreas and venom duct of C. litterantus gave the highest yield of the genomic DNA, but with poor fragment integrity and high contamination. Moreover, specific amplification products with the expected size were obtained through PCR. It is concluded that the best method to acquire the genomic DNA from C. litteratus is to extract the genomic DNA from the muscular foot with the Improved DNA Rapid Extraction Kit, with which high-quality genomic DNA can be produced and meet the requirements of subsequent research. |
| |
| Keywords: | |
| 本文献已被 CNKI 等数据库收录！ |
| 点击此处可从《热带生物学报》浏览原始摘要信息 |
|
点击此处可从《热带生物学报》下载全文 |
|
