| 引用本文: | 黄夕洋 蒋水元 李 虹 李 锋.曼地亚红豆杉RAPD反应体系与程序的优化[J].广西植物,2008,(4):451-454.[点击复制] |
| HUANG Xi-Yang, JIANG Shui-Yuan, LI Hong, LI Feng.Optimization of the RAPD reaction system and procedure of Taxus media[J].Guihaia,2008,(4):451-454.[点击复制] |
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| 摘要: |
| 以曼地亚红豆杉为研究对象,采用L16(45)正交组合实验和单因素梯度实验对MgCl2、dNTP、随机引物、Taq酶、模板DNA浓度和退火温度、循环次数等影响RAPD扩增的重要因素进行优化,以期建立最优的RAPD反应体系与程序。实验结果表明,各因素最适条件为:25 μLPCR反应体系中10×Buffer 2.5 μL,MgCl2 1.5 mmol/L,dNTP 0.2 mmol/L,随机引物0.6 μmol/L,Taq酶1.0 U,模板DNA 80 ng; 退火温度为37 ℃,循环次数为45次。 |
| 关键词: 曼地亚红豆杉 RAPD优化 反应体系和程序 正交组合 单因素 |
| DOI： |
| 分类号:Q943 |
| 基金项目: |
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| Optimization of the RAPD reaction system and procedure of Taxus media |
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HUANG Xi-Yang, JIANG Shui-Yuan, LI Hong, LI Feng
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Guangxi Institute of Botany, Guangxi Zhuang Autonomous Region and the Chinese Academy of Sciences, Guilin 541006, China
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| Abstract: |
| Regarded Taxus media as the research object,several important factors included MgCl2,dNTP,random primer,Taq polymerase,template DNA and annealing temperature,amplification were optimized by L16(45)orthogonal design experiment and single factor experiment in order to establish optimal RAPD system and procedure. The results showed that a total volume of 25 μL PCR reaction system conditioned 10×Buffer 2.5 μL,MgCl2 1.5 mmol/L,dNTP 0.2 mmol/L,random primer 0.6 μmol/L,Taq polymerase1.0 U,template DNA 80ng,and annealing temperature was 37 ℃,amplification for 45 cycles. |
| Key words: Taxus media RAPD optimization reaction system and procedure orthogonal design single factor |
