| 引用本文： |
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刘金文,沙伟,滕兆岩.采用正交设计和响应曲面设计法优化芦苇随机扩增多态DNA体系[J].广西科学,2004,11(2):148-150,160. [点击复制]
- Liu Jinwen,Sha Wei,Teng Zhaoyan.Optimal of RAPD Realtion System on Phragmites australis by Orthogonal Design and Response Surface Design[J].Guangxi Sciences,2004,11(2):148-150,160. [点击复制]
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| 摘要: |
| 采用正交设计和响应曲面设计(RSD)优化适合于芦苇[Phragmites australis(Cav.) Trin.ex steud]随机扩增多态DNA(RAPD)的反应体系,结果确定芦苇RAPD反应体系的最佳方案为:在PCR扩增程序,冷启动,94℃变性1min,36℃复性90s,72℃延伸2 min,40个循环;72℃延伸7min,4℃保存;25μl反应体系包括模板DNA 60ng、三磷酸脱氧核苷酸(dNTP)7.5mmol、Taq酶1.5u、引物5pmol。 |
| 关键词: 芦苇 RAPD 正交设计 响应曲面设计 条件优化 |
| DOI： |
| 投稿时间：2003-09-12修订日期：2004-01-29 |
| 基金项目: |
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| Optimal of RAPD Realtion System on Phragmites australis by Orthogonal Design and Response Surface Design |
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Liu Jinwen, Sha Wei, Teng Zhaoyan
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| (The Life Sci and Engineering College, Qiqihaer Univ, Qiqihaer, Heilongjiang, 161006, China) |
| Abstract: |
| The optimum reaction systems of RAPD were obtained by orthogonal design and response surface design(RSD). The RAPD reaction system of Phragmites australis were determined. For a 25 μl reaction,the optional composition included 60ng of template DNA,7.5 mmol of dNTP,1.5 u of taq DNA,5 pmol of primer The program for efficient amplification included 40 cycles of denaturation at 94℃ for 1 min,annealing at 36℃ for 1.5 min,and extension at 72℃ for 2 min(7 min for the final cycle). |
| Key words: Phragmites australis RAPD orthogonal design response surface design RAPD optimal |
