| 濒危植物沉水樟ISSR-PCR反应体系的建立与优化 |
| |
| 引用本文： | 蔡丽平,梅辉坚,陈奶莲,苗作云,吴鹏飞,邹显花,马祥庆.濒危植物沉水樟ISSR-PCR反应体系的建立与优化[J].西南林业大学学报,2013(5):40-45. |
| |
| 作者姓名： | 蔡丽平 梅辉坚 陈奶莲 苗作云 吴鹏飞 邹显花 马祥庆 |
| |
| 作者单位： | [1]福建农林大学林学院,福建福州350002 [2]抚州市林业局森林资源监测中心,江西抚州344000 [3]福建农林大学生命科学学院,福建福州350002 [4]黄河科技学院,河南郑州450063 |
| |
| 基金项目： | 国家林业局林业公益性行业科研项目（201304303）资助;高校产学合作科技重大项目（2013N5002）资助. |
| |
| 摘 要： | 为从分子水平上探讨濒危物种沉水樟种质资源的保护机制,以其基因组DNA为模版,开展ISSR-PCR反应体系的研究,对各反应因子水平、引物退火温度和循环参数进行优化,确定适于沉水樟ISSR-PCR反应的最佳体系：20 μL反应体系中各反应物的最适含量为模板75 ng、引物0.4μmol/L、dNTP 200 μmol/L、Taq酶1.25 U、Mg2＋ 1.5 mmol/L;扩增程序为：94℃预变性7 min; 94℃变性30 s,53.0～58.0℃退火45 s,72℃延伸90 s,45个循环;72℃延伸7 min,4℃保存.
|
| 关 键 词： | 沉水樟 濒危植物 ISSR PCR反应体系 |
Establishment and Optimization of ISSR-PCR System for Endangered Plant Species Cinnamomun micranthum |
| |
| Affiliation: | CAI Li-ping , MEI Hui-jian, CHEN Nai-lian , MIAO Zuo-yun, WU Peng-fei ,ZOU Xian-hua ,MAXlang-qing ( 1. College of Forestry, Fujian Agriculture and Forestry University, Fuzhou Fujian 350002, China 2. Monitoring Center of Fuzhou Forest Resources, Fuzhou Jiangxi 344000, China; 3. College of Life Science, Fujian Agriculture and Forestry University, Fuzhou Fujian 350002, Chin 4. Huanghe Science and Technology College, Zhengzhou Henan 450063, China) |
| |
| Abstract: | In order to explore the germplasm resource conservation mechanism for the endangered species Cinnamomun micranthum from the molecular level,the most appropriate ISSR-PCR reaction system with its genomic DNA was experimented by optimizing the level of each reacting factors,the primer annealing temperature and cycle parameters.The results showed that the suitable dosage of each reacting factor for C.micranthum ISSR-PCR reaction system was as follows：20 μL reaction system,including 75 ng template,0.4 μmol/L primer,200 μmol/L dNTP,1.25 U Taq enzyme,and 1.5 mmol/L Mg2 ＋.The best amplification program was as：denaturation for 7 min at 94 ℃ in advance,then denaturation at 94 ℃ for 30 s,annealing for 45 s at 53.0-58.0 ℃,extension for 90 s at 72 ℃,after 45 cycles,extension at 72 ℃ for 7 min,preservation at 4 ℃. |
| |
| Keywords: | Cinnamomun micranthum endangered plants ISSR PCR reaction system |
| 本文献已被 维普 等数据库收录！ |
|
