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中国农学通报 ›› 2010, Vol. 26 ›› Issue (16): 26-30.

• 生物技术科学 • 上一篇    下一篇

余甘子SRAP反应体系的优化

周平 许奇志 熊月明 金光 郭林榕 陈由强   

  • 收稿日期:2010-03-22 修回日期:2010-05-05 出版日期:2010-08-20 发布日期:2010-08-20
  • 基金资助:
    广西农科院科技发展基金;福建省科技计划重点项目;福建省农科院青年科技人才创新基金

Optimization of Sequence-Related Amplified Polymorphism Amplification System for Phyllanthus emblica

  • Received:2010-03-22 Revised:2010-05-05 Online:2010-08-20 Published:2010-08-20

摘要: 本文报道通过正交设计和单因素优化实验建立余甘子SRAP-PCR优化体系。实验表明PCR反应各因素(模板DNA、Mg2+、dNTPs、引物)对扩增结果均有不同的影响。采用50 ng模板DNA,2.0 mmol?L-1 Mg2+, 0.4 mmol?L-1 dNTPs, 0.3 μmol?L-1 引物的20 μL反应体系可扩增出最清晰丰富的多样性条带。使用该优化体系检测12份余甘子种质资源样品,结果稳定可信,适用于分子遗传研究。

关键词: 麻地膜, 麻地膜, 土壤pH值, 微生物

Abstract: In this paper, a optimized SRAP-PCR amplification system for Phyllanthus emblica was established by the orthogonal design and single factor optimization. It suggested that each factor (template DNA、Mg2+、dNTPs、primers)had different effect on the results of PCR, and 50 ng template DNA, 2.0 mmol/L-1 Mg2+, 0.4 mmol/L-1 dNTPs, 0.3 μmol/L-1 primers in a total of 20 μL reaction solution could be able to amplified the most rich polymorphism and clear bands. The optimized SRAP-PCR system was tested on twelve Phyllanthus emblica germplasms and shown to be steady and reliable for molecular genetics research.