GUO Jing-liang,RUAN Cheng-jiang.DNA Extraction of Xanthoceras sorbifolia Bunge and Optimization of RAPD and ISSR Reaction Systems[J].HEILONGJIANG AGRICULTURAL SCIENCES,2015,(10):27-29,30.[doi:10.11942/j.issn1002-2767.2015.10.0027]
文冠果DNA提取及RAPD和ISSR反应体系构建
- Title:
- DNA Extraction of Xanthoceras sorbifolia Bunge and Optimization of RAPD and ISSR Reaction Systems
- 文章编号:
- 1002-2767（2015）10-0027-03
- Keywords:
- Xanthoceras sorbifolia; DNA; RAPD; ISSR
- 分类号:
- S794
- 文献标志码:
- A
- 摘要:
- 为研究文冠果种质遗传多样性，以新疆奇台文冠果自然栽培居群的48个单株为材料，采用改良CTAB法提取文冠果基因组DNA，并对其RAPD和ISSR反应体系进行了优化。研究结果表明：文冠果的RAPD最适反应体系为：PCR扩增总体积为20 μL，包含30 ng的模板DNA，10×PCR buffer 2 μL，2.0 mmol?L-1 Mg 2+，0.1 mmol?L-1dNTP和Taq DNA聚合酶1 U；ISSR最适反应体系为：PCR扩增总体积为20 μL，包括30 ng的模板DNA，10×PCR buffer 2 μL，2.5 mmol?L-1 Mg2+， 0.2 mmol?L-1dNTP和Taq DNA聚合酶1 U。在最适反应体系下，RAPD和ISSR分析具有良好的稳定性和可重复性。
- Abstract:
- In order to study the genetic diversity of Xanthoceras Sorbifolia germplasms,taking 48 individuals as materials growing in natural cultivated population in the Qitai county of Xinjiang germpalsm nursery.Leaves DNA of each sample were extracted using the modified CTAB method,RAPD and ISSR reaction systems were optimized.The results showed that the optimal RAPD reaction system was in the total volume of 20 μL for PCR amplification,containing 30 ng template DNA,2 μL 10 × PCR buffer,2.0 mmol?L-1 Mg2+，0.1 mmol?L-1 dNTPs,1 U Taq enzyme.The optimal ISSR analysis reaction system was in the total volume of 20 μL for PCR amplification,containing 30 ng template DNA,2 μL 10 × PCR buffer,2.5 mmol?L-1 Mg2+，0.2 mmol?L-1 dNTPs,1 U Taq enzyme.In these optimum reaction systems,RAPD and ISSR analysis had stability and repeatability very well.
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备注/Memo
收稿日期：2015-08-28
基金项目：国家“十二五”科技支撑计划专题资助项目(2015BAD07B01)；辽宁省自然科学基金资助项目(2014020135)；辽宁省大学生创新创业训练计划资助项目(S201412026000022)
第一作者简介：郭井亮(1992-)，男，辽宁省沈阳市人，学士，从事资源植物分子育种研究。E-mail:461738609@qq.com。
通讯作者：阮成江(1972-)，男，河南省信阳市人，博士，教授，博士研究生导师，从事油料能源植物种质创新与开发利用研究。E-mail：ruan@dlnu.edu.cn。