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中国农学通报 ›› 2008, Vol. 24 ›› Issue (9): 457-460.

所属专题: 马铃薯

• 农学 农业基础科学 • 上一篇    下一篇

木薯SRAP扩增体系的建立与优化

夏志强,邹枚伶,王文泉   

  • 收稿日期:2008-06-11 修回日期:2008-07-03 出版日期:2008-09-05 发布日期:2008-09-05

Optimization of SRAP Reaction System in Cassava

Xia Zhiqiang, Zou Meiling, Wang Wenquan   

  • Received:2008-06-11 Revised:2008-07-03 Online:2008-09-05 Published:2008-09-05

摘要: 建立适宜木薯DNA的SRAP扩增体系,为木薯分子标记和基因图谱的构建打下基础。以木薯基因组DNA为模板,采用序列相关扩增多态性(sequence related amplified polymorphism,SRAP)技术对木薯DNA进行PCR扩增,逐级优化反应参数。最佳SRAP-PCR反应体系(10Ll)为:DNA (50ng/μl) 0.5μl、10×PCR buffer (Mg2+) 1.0μl、dNTPs (20mM) 0.2μl、primer (50ng) 0.3μl、Taq polymerase (5U/μl) 0.2μl。该程序和体系能很好地满足木薯基因组SRAP扩增的要求,SRAP标记能够很好应用于木薯遗传研究。

Abstract: The aim of the research was to establish SRAP amplification system which was suitable for Cassava DNA so as to lay foundation for molecular marker and the construction of gene map in Cassava. Using Cassava genome DNA as template, sequence-related amplified polymorphism (SRAP) was applied for Cassava to carry on PCR amplification of its DNA and optimize the reaction parameter grade by grade. The stable and reproducible SRAP reaction system of Cassava has been developed. The optimum SRAP PCR system (total of 10μl) was as follows: DNA (50ng/μl) 0.5μl, 10×PCR buffer (Mg2+) 1.0μl, dNTPs (20mM) 0.2μl, primer (50ng) 0.3μl, Taq polymerase (5U/μl) 0.2μl. The program and system could meet the demands for genome SRAP amplification in Cassava. It was feasible to apply SRAP marker in genetic research in Cassava.

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