• 中文核心期刊
  • CSCD来源期刊
  • 中国科技核心期刊
  • CA、CABI、ZR收录期刊

空心菜SRAP-PCR分析体系的建立与优化

Establishment and Optimization of SRAP Amplification System in Lpomoea aquatica

  • 摘要: 以空心菜为试验材料,利用单因素试验对影响SRAP-PCR反应体系的DNA模板、10×PCR Buffer(含Mg2+)、dNTPs、引物以及TaqDNA聚合酶5个因素进行优化,从而建立适合于空心菜SRAP-PCR分析的反应体系。结果表明:空心菜SRAP-PCR最佳的反应体系(25μL)为:DNA模板80ng、10×PCR Buffer(含Mg2+)2.5mmol·L-1、dNTPs 0.2mmol·L-1、引物0.4μmol·L-1、TaqDNA聚合酶1U。运用所优化的SRAP-PCR分析体系,对“本地空心菜”和“三叉空心菜”2个品种进行SRAP-PCR扩增,扩增结果所获得的电泳谱带清晰明亮,材料间表现出明显的多态性,并且在25对SRAP引物组合中可以筛选出7对多态性高的组合,能明显区分2个空心菜品种。由此可见,本试验所建立的SRAP-PCR体系适用于空心菜的品种鉴定和遗传多样性分析。

     

    Abstract: To establish the optimal sequence-related amplified polymorphism(SRAP-PCR)system of Lpomoea aquatica,the concentrations of template DNA,10×PCR Buffer(including Mg2+),dNTPs,primer and Taq DNA polymerase,which maybe affect the SRAP-PCR reactions,were optimized by single factor test.The results showed that the optimal SRAP amplification system of lpomoea aquatica was established,which containing 80 ng template DNA,2.5mmol·L-1 10×PCR Buffer(including Mg2+),0.2mmol·L-1 dNTPs,0.4μmol·L-1primer and 1U Taq DNA polymerase.The molecular identification of two Lpomoea aquatica cultivars(‘Ben di'and ‘San cha')were conducted with the optimized SRAP marker system.7out of 25 SRAP primer combinations were selected to distinguish two Lpomoea aquatica cultivars and the amplification products were clear,bright and showed abundant polymorphism bands, which indicated that the optimized SRAP-PCR system could be applied in cultivars identification and genetic diversity research of Lpomoea aquatica.

     

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