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| 红松SSR-PCR反应体系的建立与优化 | |
| 引用本文： | 冯富娟,赵丹,孙晓艳,韩士杰,许小妍.红松SSR-PCR反应体系的建立与优化[J].经济林研究,2010,28(1):35-40. |
| 作者姓名： | 冯富娟 赵丹 孙晓艳 韩士杰 许小妍 |
| 作者单位： | 1. 东北林业大学生命科学学院,黑龙江哈尔滨,150040 2. 东北林业大学林学院,黑龙江哈尔滨,150040 3. 中国科学院沈阳应用生态研究所,辽宁沈阳,110016 |
| 基金项目： | 中科院知识创新工程重要方向项目(SCX2-YW-N-031-2);;黑龙江省博士后科研启动金资助 |
| 摘 要： | 为了建立红松SSR-PCR最佳反应体系,以CTAB法提取的红松针叶DNA为模板,应用单因子试验及L16(45)正交试验系统分析了DNA模板浓度、Mg2+浓度、dNTPs浓度、引物浓度、Taq酶浓度等对SSR-PCR扩增结果的影响,并对延伸时间、热循环参数进行了探讨,建立了稳定的、可重复的红松SSR-PCR最佳反应体系及PCR扩增参数。结果表明,PCR的最佳反应体系为:10μL体系中,1×PCRbuffer1μL、DNA模板50～120ng,Mg2+3mmol/L,dNTP0.4mmol/L,引物0.5μmol/L,Taq酶0.75U。利用该体系筛选出扩增条带清晰、多态性丰富的SSR引物13对。在此反应体系下,所扩增谱带清晰、稳定、多态性高。 |
| 关 键 词： | 红松 SSR 优化 正交设计 单因子试验 |
Establishment and optimization of the SSR-PCR reaction system in Pinus koraiensis Sieb.et Zucc. |
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| FENG Fu-juan,ZHAO Dan,SUN Xiao-yan,HAN Shi-jie,XU Xiao-yan.Establishment and optimization of the SSR-PCR reaction system in Pinus koraiensis Sieb.et Zucc.[J].Economic Forest Researches,2010,28(1):35-40. | |
| Authors: | FENG Fu-juan ZHAO Dan SUN Xiao-yan HAN Shi-jie XU Xiao-yan |
| Institution: | 1. a. School of life science;b. School of Forestry;Northeast Forestry University;Harbin 150040;Heilongjiang;China;2.Institute of Applied Ecology;Chinese Academy of Sciences;Shenyang 110016;Liaoning;China |
| Abstract: | In order to establish the optimal SSR-PCR reaction system in Pinus koraiensis, taking DNA extracted from needle as template, the effects of concentrations of template DNA, Mg2+, dNTPs, primer and Taq polymerase on SSR-PCR amplification were analyzed by the method of CTAB using single factor test and orthogonal test of L16(45), and the extension time and the parameters of thermal cycling were researched. The optimal SSR-PCR reaction system and the amplification parameters were established, and the system was... |
| Keywords: | Pinus koraiensis SSR optimization orthogonal design single factor test |
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