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中国农学通报

中国农学通报 ›› 2010, Vol. 26 ›› Issue (15): 86-88.

• 生物技术科学 • 上一篇    下一篇

橄榄SRAP-PCR体系的建立和优化

张平湖 刘冠明   

  • 收稿日期:2010-02-20 修回日期:2010-03-15 出版日期:2010-08-05 发布日期:2010-08-05
  • 基金资助:

    广东省农业科技计划项目

Establishment and Optimization of SRAP-PCR System in Canarium album Reausch

  • Received:2010-02-20 Revised:2010-03-15 Online:2010-08-05 Published:2010-08-05

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摘要：

以橄榄品种为材料，采用L16（45）的正交试验设计，对影响PCR反应的Taq酶量、Mg2+浓度、模板DNA含量、dNTPs浓度和引物浓度5个因素进行了SRAP-PCR扩增反应条件优化研究，并利用反应体系对11个橄榄品种进行了SRAP-PCR扩增。结果表明：在20μl体系中，Taq酶1.5U、Mg2+2.5 mmol/L、模板DNA 60ng、dNTPs 0.2 mmol/L和引物0.15μmol/L时的扩增效果最好；利用该体系，SRAP标记引物对Me5- Em2在11个橄榄品种中可以扩增出7条清晰的多态性条带。

关键词: 加拿大, 加拿大, 禽流感, 补偿政策, 启示

Abstract:

the variety of Canarium album Reausch was used as material, Optimization of SRAP-PCR system was studied with L16（45）orthogonal design of five factors (Taq polymerase, Mg2+, DNA template, dNTPs and primer) and the DNA of 11 varieties were amplified with the optimized SRAP-PCR system. The results showed that in 20μl amplification system contained 1.5U Taq DNA polymerase, 2.5mmol/L Mg2+, 60ng genomic templates DNA, 0.2mol/L dNTPs, and 0.15μmol/L Primer, the result of SRAP-PCR was best for Canarium album Reausch. Seven polymorphic bands were amplified with one primer pair Me5-Em2 through the optimized SRAP-PCR system in 11 varieties of Canarium album Reausch.