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中国农学通报

中国农学通报 ›› 2009, Vol. 25 ›› Issue (4): 30-34.

所属专题： 园艺

• 生物技术科学 • 上一篇    下一篇

丝瓜SRAP-PCR体系建立与优化

吴 红,林 清,雷开荣,陈 旭,蒋晓英,陶伟林   

  • 收稿日期:2008-12-01 修回日期:2009-01-07 出版日期:2009-02-20 发布日期:2009-02-20

Establishment and Optimization of SRAP-PCR Amplification System in loofah

  • Received:2008-12-01 Revised:2009-01-07 Online:2009-02-20 Published:2009-02-20

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摘要： 本文以丝瓜(Luffa cylindrica (L.) M. Roem)基因组DNA为模板，采用正交试验设计，对SRAP (sequence related amplified polymorphism, 序列相关扩增多态性)反应体系中的4种关键因素（dNTPs、Taq酶、引物、模板）进行优化，建立了适合于丝瓜基因组SRAP分子标记的扩增体系：反应总体积10μl，其中含Taq酶1.0 U、模板DNA 50.00 ng、dNTPs 0.3mmol/L、引物0.30μmol/L。该体系能很好地满足丝瓜基因组SRAP标记的要求，可应用于丝瓜的分子生物学研究。

关键词: 冬小麦, 冬小麦, PEG-6000模拟干旱, 水势, 叶绿素

Abstract: In this study, orthogonal experiments were designed to optimize the key parameters (dNTPs、Taq polymerase、primer、template DNA) of SRAP-PCR amplification system in loofah（Luffa cylindrica (L.) M. Roem）. The results showed that the optimized reaction system included: DNA template 50ng, dNTPs 0.3mmol/L, primer 0.30μmol/L and Taq polymerase 1U in the 10ul total volume. This system is fit for the genome amplification of loofah and can be well used for the molecular biological research in loofah.

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