| 利用正交设计优化甘蔗SRAP-PCR反应体系 |
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| 引用本文： | 昝逢刚,刘家勇,赵俊,赵培方,吴转娣,吴才文.利用正交设计优化甘蔗SRAP-PCR反应体系[J].中国甜菜,2011(3):6-8. |
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| 作者姓名： | 昝逢刚 刘家勇 赵俊 赵培方 吴转娣 吴才文 |
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| 作者单位： | 云南省农业科学院甘蔗研究所／云南省甘蔗遗传改良重点实验室,开远616600 |
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| 基金项目： | 现代农业产业技术体系建设专项资金（nycytx-024-01-03）; 云南省“十一五”科技攻关（2006NG11） |
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| 摘 要： | 利用正交设计L16（45）对甘蔗SRAP-PCR反应体系的五大因素（Mg2＋、dNTPs、引物、模板DNA、Taq酶）在4个水平上进行优化,得到如下结论：各因素水平变化对PCR反应的影响从大到小依次是：Mg^2＋、dNTPs、引物、Taq酶和模板DNA;通过对各因素进行筛选,建立甘蔗SRAP-PCR反应的最佳体系（20μL）为：dNTPs 0.25 mmol/L、引物0.1μmol/L、Mg^2＋2.5 mmol/L、Taq酶0.25U和模板DNA 60 ng。
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| 关 键 词： | 甘蔗 SRAP-PCR 正交设计 优化 |
Optimization for Sugarcane SRAP-PCR System by Orthogonal Design |
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| Authors: | ZAN Feng-gang LIU Jia-yong ZHAO Jun ZHAO Pei-fang WU Zhuan-di WU Cai-wen |
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| Institution: | (Yunnan Province Key Laboratory of Sugarcane Genetic Improvement /Sugarcane Research Institute,Yunnan Academy of Agricultural Sciences,Kuaiyuan 616600,China) |
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| Abstract: | The orthogonal design was used to optimize SRAP-PCR amplification system on sugarcane in four levels of five factor（Mg2＋,dNTPs,Taq DNA polymerase,primer and the DNA template）,respectively.The result of PCR showed that the effect of each factor in different levels in the order from big to small was Mg2＋,dNTPs,primer,Taq DNA polymerase and the DNA template.A most suitable SRAP-PCR system for sugarcane was established,the system of 20μl reaction contained 0.25 mmol/L dNTPs,0.1μmol/L primer,2.5 mmol/L Mg2＋,0.25U Taq DNA polymerase and 60 ng DNA template. |
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| Keywords: | sugarcane SRAP orthogonal design optimization |
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