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中国农学通报

中国农学通报 ›› 2008, Vol. 24 ›› Issue (4): 37-41.

所属专题： 油料作物

• 生物技术科学 • 上一篇    下一篇

花生SSR-PCR体系的优化

张 发,万勇善,刘风珍   

  • 收稿日期:2008-01-29 修回日期:2008-02-20 出版日期:2008-04-05 发布日期:2008-04-05

Optimization of SSR – PCR Reaction System of Peanut

Zhang Fa, Wan Yongshan, Liu Fengzhen   

  • Received:2008-01-29 Revised:2008-02-20 Online:2008-04-05 Published:2008-04-05

PDF (PC)

1088

摘要： 以花生品种丰花3号为材料，研究了花生SSR技术中PCR反应体系的主要成分对SSR 扩增效果的影响及不同引物的退火温度。结果表明dNTP 对扩增影响较大，每对引物都有其扩增适合的退火温度。确立了适合花生SSR 分子标记研究的优化体系。最终确定总反应体系为20 μl，其中25 mM MgCl2 1.5 μl，2.5 mM dNTP 1.6 μl，5 U/μl Taq 酶 0.17 μl，100 ng/μl模板DNA 0.4 μl，2.5 mM 引物 5 μl。优化后的扩增程序退火温度为54℃。

关键词: 微肥, 微肥, 营养, 紫花苜蓿

Abstract: FengHua 3 was used in this study to optimize the several factors applied in SSR technique system. dNTP was found to have distinct effect to amplification, and annealing temperature specific to primer pair was also checked. Optimized SSR system for peanut was established : 25 mM MgCl2 1.5 μl, 2.5 mM dNTP 1.6 μl, 5 U/μl Taq 0.17 μl, 100 ng/μl template DNA 0.4 μl, 2.5 mΜ primers 5 μl were added to the PCR mixture. Optimized annealing temperature was 54℃.