| 樟树ISSR-PCR反应体系优化研究 |
| |
| 引用本文： | 邢建宏,陈存及,张国防,彭东辉,刘宝,陈碧华.樟树ISSR-PCR反应体系优化研究[J].福建林业科技,2006,33(3):96-100. |
| |
| 作者姓名： | 邢建宏 陈存及 张国防 彭东辉 刘宝 陈碧华 |
| |
| 作者单位： | 1. 福建农林大学林学院,福建,福州,350002
2. 福建省林业科学研究院,福建,福州,350012 |
| |
| 摘 要： | 以樟树基因组DNA为材料,分析了影响樟树ISSR-PCR的主要参数,确立了适用于樟树的ISSR反应体系和扩增条件。结果表明获得清晰、重复性高的樟树ISSR-PCR扩增条件为:20μL PCR反应体积中,1×PCR Buffer(10 mmol.L-1Tris.HCl,pH8.3,50 mmo.lL-1KCl),2.0 mmo.lL-1MgCl2,200μmol.L-1dNTPs,50 ng模板DNA,200 nmo.lL-1引物,0.5 UTaq DNA聚合酶。在扩增过程中,引物的适宜退火温度比其Tm值平均高3℃。这为今后利用ISSR标记技术开展樟树种间遗传多样性分析提供参考。
|
| 关 键 词： | 樟树 ISSR PCR反应体系优化 |
| 文章编号： | 1002-7351（2006）03-0096-05 |
| 收稿时间： | 2006-02-14 |
| 修稿时间： | 2006-02-142006-04-29 |
Study on the Establishment of ISSR-PCR Reaction Conditions in Cinnamomum camphora |
| |
| XING Jian-hong,CHEN Cun-ji,ZHANG Guo-fang,PENG Dong-hui,LIU Bao,CHEN Bi-hua.Study on the Establishment of ISSR-PCR Reaction Conditions in Cinnamomum camphora[J].Journal of Fujian Forestry Science and Technology,2006,33(3):96-100. |
| |
| Authors: | XING Jian-hong CHEN Cun-ji ZHANG Guo-fang PENG Dong-hui LIU Bao CHEN Bi-hua |
| |
| Abstract: | In this paper,the main parameters of affecting the result of ISSR-PCR amplification was analyzed using the materials of Cinnamomum camphora DNA and the optimal ISSR-PCR reaction condition in Cinnamomum camphora was established.The results showed that the optimum concentration of five important components i.e.MgCl_2,dNTPs,template DNA,primer,Taq DNA polymerase in 20 μL reaction system of ISSR was 2.0 mmol·L~-1,200 μmol·L~1,50 ng,200 nmol·L~-1 and 0.5 U,respectively.The appropriate annealing temperature was average 3℃ higher than Tm of primer.The results provided a reference for the analysis of inter-species genetic diversity of Cinnamomum camphora. |
| |
| Keywords: | Cinnamomum camphora(L )Presl ISSR PCR reaction condition |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
|
