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王家保, 刘志媛, 徐碧玉, 邓穗生, 杜中军, 陈业渊. 用正交设计优化荔枝RAPD反应体系[J]. 植物科学学报, 2005, 23(4): 363-368.
引用本文: 王家保, 刘志媛, 徐碧玉, 邓穗生, 杜中军, 陈业渊. 用正交设计优化荔枝RAPD反应体系[J]. 植物科学学报, 2005, 23(4): 363-368.
WANG Jia-Bao, LIU Zhi-Yuan, XU Bi-Yu, DENG Sui-Sheng, DU Zhong-Jun, CHEN Ye-Yuan. Optimization of the RAPD Reaction System of Litchi chinensis Sonn. by Orthogonal Design[J]. Plant Science Journal, 2005, 23(4): 363-368.
Citation: WANG Jia-Bao, LIU Zhi-Yuan, XU Bi-Yu, DENG Sui-Sheng, DU Zhong-Jun, CHEN Ye-Yuan. Optimization of the RAPD Reaction System of Litchi chinensis Sonn. by Orthogonal Design[J]. Plant Science Journal, 2005, 23(4): 363-368.

用正交设计优化荔枝RAPD反应体系

Optimization of the RAPD Reaction System of Litchi chinensis Sonn. by Orthogonal Design

  • 摘要: 以改良CTAB法提取的荔枝基因组DNA为模板,应用L25(56)正交表,研究了Taq、Mg2+、随机引物、dNTPs和DNA模板5种RAPD反应组分浓度变化对扩增结果的影响,量化分析结果表明:正交设计可以应用于RAPD反应体系的建立。用这种方法建立的荔枝RAPD-PCR优化反应体系为:25μL反应体系中含1×Buffer、2.0mmol/LMg2+、2.0UTaqDNA聚合酶、0.15mmol/LdNTPs、0.6μmol/L随机引物、25ngDNA模板。

     

    Abstract: The L25(56) orthogonal diagram was applied to study the effects of different concentrations of Taq polymerase, Mg2+, dNTPs, primer and DNA on litchi (Litchi chinensis Sonn.) RAPD-PCR results and to get a suitable PCR system. The results showed that the orthogonal design could be used to optimize the RAPD-PCR system. A suitable RAPD reaction system for litchi was developed, i.e.1×Taq polymerase reaction buffer, 2.0 mmol/L Mg2+, 2.0 U Taq DNA polymerase, 0.15 mmol/L dNTPs, 0.6 μmol / L primer and 25 ng genomic DNA templates with total 25 μL reaction solution.

     

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