| 石蒜属植物SCoT-PCR反应体系构建及优化 |
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| 引用本文： | 姜小凤,高燕会,童再康,黄春红.石蒜属植物SCoT-PCR反应体系构建及优化[J].浙江农林大学学报,2013,30(3):444-452. |
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| 作者姓名： | 姜小凤 高燕会 童再康 黄春红 |
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| 作者单位： | 1.浙江农林大学 亚热带森林培育国家重点实验室培育基地，浙江 临安 311300 |
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| 摘 要： | 建立并优化了石蒜属Lycoris植物的目标起始密码子多态性-聚合酶链式反应（SCoT-PCR）反应体系，为研究石蒜属种质资源的遗传多样性、亲缘关系及遗传改良提供新的思路。采用L25（56）正交试验和单因素试验2种方法优化石蒜属植物SCoT-PCR体系。得出石蒜属植物SCoT-PCR 最佳反应体系:DNA 模板2.0 mgL－1，引物0.125 molL－1，三磷酸碱基脱氧核苷酸（dNTPs ）0.2 mmolL－1，镁离子（Mg2+） 3.0 mmolL－1，Taq DNA聚合酶1.016.67 nkat。对优化的反应体系进行通用性、稳定性及可靠性检测，结果均能获得丰富、稳定、清晰的DNA谱带。最后可知SCoT 新型标记在石蒜属植物中应用效果良好，可为石蒜属植物以后的研究提供基础条件。图7表6参25
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| 关 键 词： | 植物学 石蒜属 目标起始密码子多态性 PCR反应体系 正交设计 单因素试验 |
| 收稿时间： | 2012-05-02 |
Establishing and optimizing a SCoT-PCR system for Lycoris |
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| JIANG Xiaofeng,GAO Yanhui,TONG Zaikang,HUANG Chunhong.Establishing and optimizing a SCoT-PCR system for Lycoris[J].Journal of Zhejiang A&F University,2013,30(3):444-452. |
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| Authors: | JIANG Xiaofeng GAO Yanhui TONG Zaikang HUANG Chunhong |
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| Institution: | 1.The Nurturing Station for the State Key Laboratory of Subtropical Silviculture, Zhejiang A & F University, Lin’an 311300, Zhejiang, China |
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| Abstract: | An optimal SCoT（start condon tagreted polymorphism）-PCR（polymerase chain reaction）amplification system was established to provide a new way of studying genetic diversity and genetic improvement in Lycoris. This system was optimized based on the L25（56）orthogonal experiment and single factor experiment. Results showed that the optimal reaction system （20 L）contained 2.0 mgL－1 DNA template, 0.125 molL－1 primers, 0.2 mmolL－1 dNTPs, 3.0 mmolL－1 Mg2+, and 1.0 16.67 nkat Taq DNA polymerase. 34 materials of Lycoris were randomly selected and the test was repeated 3 times. Using this optimal reaction system, stable and clear polymorphic bands were acquired. These favorable results could provide a scientific basis for further study of Lycoris.［Ch, 7 fig. 6 tab. 25 ref.］ |
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