| 引用本文: | 郑道君, 谢良商, 张 文, 张治礼.海南龙血树RAPD-PCR反应体系的优化[J].广西植物,2011,(1):31-35.[点击复制] |
| ZHENG Dao-Jun, XIE Liang-Shang, ZHANG Wen, ZHANG Zhi-Li.Optimization of RAPD-PCR reaction systemfor Dracaena cambodiana[J].Guihaia,2011,(1):31-35.[点击复制] |
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| 摘要: |
| 采用单因子试验和正交试验设计,对影响海南龙血树RAPD-PCR反应的模板DNA量、Mg2+、dNTP和引物浓度,Taq聚合酶用量等因子进行研究,分析各因子对RAPD-PCR扩增结果的影响,确立了海南龙血树RAPD-PCR反应的最佳条件,即在25 μL反应体系中,包含10×buffer 2.5 μL,2.0 mmol/L MgCl2,300 μmol/L dNTPs,Taq酶1.5U,引物0.8 μmol/L和DNA模板20 ng。以此条件为基础,筛选出适用于海南龙血树RAPD-PCR分析的18条有效引物,为采用RAPD技术分析海南龙血树种群遗传变异水平和遗传结构打下了基础。 |
| 关键词: 海南龙血树 RAPD 反应条件优化 有效引物筛选 |
| DOI：10.3969/j.issn.1000-3142.2011.01.007 |
| 分类号:Q943 |
| 基金项目: |
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| Optimization of RAPD-PCR reaction systemfor Dracaena cambodiana |
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ZHENG Dao-Jun, XIE Liang-Shang, ZHANG Wen, ZHANG Zhi-Li*
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Key Laboratory of Crop Genetics and Breeding, Hainan Academy of Agricultural Sciences, Haikou 571100, China
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| Abstract: |
| Both single factor test and orthogonal design were applied to study the effects of main factors on the RAPD-PCR system for Dracaena cambodiana,in which the main factors included the content of template DNA,the concentration of Mg2+、dNTPs and primers and the content of Taq DNA polymerase,and an optimal 25μL RAPD-PCR reaction system for D.cambodiana was established,including 2.5 μL 10×PCR buffer,2.0 mmol/L MgCl2,300 μmol/L dNTPs,1.5 U Taq polymerase,0.8 μmol/L primers,and 20 ng DNA template. 18 primers were selected for RAPD-PCR analysis for D.cambodiana using the optimized amplification system above,which can be employed for the analysis of genetic variation and structure of D.cambodiana population. |
| Key words: Dracaena cambodiana RAPD optimization of reaction condition screening of effective primers |
