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中国农学通报 ›› 2009, Vol. 25 ›› Issue (4): 24-29.

• 生物技术科学 • 上一篇    下一篇

椰子ISSR体系优化

潘坤,王文泉,吴翼, 唐龙祥   

  • 收稿日期:2008-11-19 修回日期:2008-12-23 出版日期:2009-02-20 发布日期:2009-02-20

Optimization of ISSR-PCR system in coconut(Cocos nucifera L.)

  • Received:2008-11-19 Revised:2008-12-23 Online:2009-02-20 Published:2009-02-20

摘要: 本实验旨在获得最佳的椰子ISSR-PCR反应体系,以海南本地高种和马来西亚高种为实验材料,采用单因素实验法将反应体系的五个主要因素设定五个梯度,根据每个因素量的变化对扩增结果产生的影响进行了研究;并利用梯度PCR仪得出引物807的最适退火温度为56.0℃;最后确定最佳反应体系为:总体积20μL,Taq聚合酶1.0U、Mg2+浓度即10×Taq Buffer用量2.5mmol/L、模板DNA用量50ng、dNTPs浓度0.2mmol/L、引物浓度0.8μmol/L。

关键词: 烟草, 烟草, 氯, 硫, 对应分析, 化学元素

Abstract: The coconut ISSR-PCR reaction system was optimized, we distilled the total DNA of two coconut cultivars that were Hainan Tall and Malaysia Tall, and the single factor experiment was designed which included five factors with five different concentrations and then the effect of the variations of these concentrations were studied. Meanwhile, we obtained the annealing temperature of primer 807, the result shows the optimal annealing temperature was 56℃, at last, the optimum reaction system for coconut consists of 0.5~2.0U Taq DNA polymerse, 2.5~3.0mmol/L 1 0×Taq Buffer(25mmol/L Mg2+), 50ng template DNA, 0.2~0.5mmol/L dNTPs and 0.8~1.6μmol/L primer with total 20μL reaction solution.

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