| 引用本文： |
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史艳财,邹蓉,朱成豪,熊忠臣,蒋运生.极小种群喙核桃ISSR-PCR反应条件的建立与优化[J].广西科学院学报,2020,36(1):78-82. [点击复制]
- SHI Yancai,ZOU Rong,ZHU Chenghao,XIONG Zhongchen,JIANG Yunsheng.Establishment and Optimization of the ISSR-PCR Reaction System for Annamocarya sinensis[J].Journal of Guangxi Academy of Sciences,2020,36(1):78-82. [点击复制]
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| 摘要: |
| 本文采用正交和单因素相结合的方法研究dNTPs、Mg2+、引物、Taq DNA聚合酶、模板DNA、循环次数及退火温度对喙核桃ISSR-PCR扩增效果的影响，以期为喙核桃遗传多样性评价、亲缘关系分析等研究奠定基础。结果表明：喙核桃最佳ISSR-PCR扩增体系及程序为20 μL的反应体系含dNTPs 0.5 mmol/L、Mg2+ 3.0 mmol/L、引物0.8 μmol/L、Taq DNA聚合酶2.5 U、模板DNA 45 ng、10×PCR Buffer 2.0 μL。退火温度为54.4℃，循环次数为40。本研究建立的喙核桃ISSR-PCR反应体系稳定、可靠，可用于该物种遗传多样性分析。 |
| 关键词: 极小种群 喙核桃 ISSR-PCR 正交设计 单因素试验 |
| DOI：10.13657/j.cnki.gxkxyxb.20200317.011 |
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| 基金项目:国家自然科学基金项目（31960276）和广西植物研究所基本业务费项目（桂植业19004）资助。 |
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| Establishment and Optimization of the ISSR-PCR Reaction System for Annamocarya sinensis |
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SHI Yancai1, ZOU Rong1, ZHU Chenghao2, XIONG Zhongchen1, JIANG Yunsheng1
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| (1.Guangxi Institute of Botany, Guangxi Zhuang Autonomous Region and Chinese Academy of Sciences, Guilin, Guangxi, 541006, China;2.College of Pharmacy, Guilin Medical University, Guilin, Guangxi, 541004, China) |
| Abstract: |
| This article used a combination of orthogonal and single-factor methods to study the effects of dNTPs,Mg2+,primers,Taq DNA polymerase,template DNA,number of cycles,and annealing temperature on the ISSR-PCR amplification effect of Annamocarya sinensis.It is hoped to lay the foundation for the research on genetic diversity evaluation and genetic relationship analysis of Annamocarya sinensis.The results showed that the optimal ISSR-PCR amplification system and procedure for Annamocarya sinensis was a 20 μL reaction system,which contained dNTPs 0.5 mmol/L,Mg2+ 3.0 mmol / L,primers 0.8 μmol/L,Taq DNA polymerase 2.5 U,template DNA 45 ng,and 10×PCR buffer 2.0 μL.The annealing temperature was 54.4℃,and the number of cycles was 40.The ISSR-PCR reaction system established in this study is stable and reliable,which can be used for genetic diversity analysis of A.sinensis. |
| Key words: minimal population Annamocarya sinensis ISSR-PCR orthogonal design single factor test |
