| 甘草RAPD-PCR反应体系正交优化研究 |
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| 引用本文： | 张增福,董建力,李明.甘草RAPD-PCR反应体系正交优化研究[J].安徽农业科学,2011,39(29):17788-17789,17818. |
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| 作者姓名： | 张增福 董建力 李明 |
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| 作者单位： | 宁夏农业技术推广总站,宁夏银川,750001;宁夏农林科学院农业生物技术研究中心,宁夏银川,750002;宁夏农林科学院沙漠化治理研究所,宁夏银川,750002 |
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| 基金项目： | 国家“十一五”支撑项目(2006BAI06A15-11) |
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| 摘 要： | 目的]建立一套适合甘草分子学研究的RAPD-PCR反应体系。方法]以甘草种质为试材,采用正交试验法设计,对影响RAPD-PCR扩增的主要因素dNTPs、引物、Taq酶和DNA模板进行优化筛选。结果]总体积25μl的甘草RAPD-PCR最佳反应体系为：10×PCR缓冲液（含MgCl2）2.5μl,10 mmol/L dNTPs 2.5μl,50 ng DNA 2.0μl,10μmol/L引物2.0μl,5 U/μl Taq酶0.4μl。对引物的退火温度进行了梯度筛选,34℃时扩增效果较好。结论]进行甘草RAPD-PCR反应体系的正交优化非常有效。
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| 关 键 词： | 甘草 DNA RAPD-PCR 反应体系 优化 正交试验 |
Optimization of RAPD-PCR Reaction System for Glycyrrhiza uralensis Based on Orthogonal Design |
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| Institution: | ZHANG Zeng-fu et al(Crop Institute of Ningxia Academy of Agriculture and Forestry Sciences,Yinchuan,Ningxia 750001) |
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| Abstract: | Objective] The aim was to obtain the optimum RAPD-PCR reaction system for Glycyrrhiza uralensis.Method] Orthogonal design was adopted to screen the suitable concentration of major factors(dNTPs,primer,Taq polymerase,DNA template) in PCR reaction system.Result] The optimal protocol was accomplished by orthogonal design in 25 μl reaction volume containing 10×PCR buffer solution(include MgCl2) 2.5 μl,10 mmol/L dNTPs 2.5 μl,50 ng DNA template 2.0 μl,10 μmol/L primer 2.0 μl,5 U/μl Taq polymerase 0.4 μl;the optimum annealing temperature was 34 ℃.Conclusion] Orthogonal design was an effective method for the optimization of RAPD-PCR reaction system for G.uralensis. |
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| Keywords: | Glycyrrhiza uralensis DNA RAPD-PCR Reaction system Optimization Orthogonal design |
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