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作 者 ：王家保;刘志媛;徐碧玉;邓穗生;杜中军;陈业渊

期 刊 ：武汉植物学研究 2005年 23卷 4期 页码：363-368

关键词：荔枝；RAPD；反应体系；正交设计；

Keywords:Litchi, RAPD, Reaction system, 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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