| 仙茅随机扩增多态性DNA反应体系的优化 |
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| 引用本文： | 李隆云,陈大霞,李泉森. 仙茅随机扩增多态性DNA反应体系的优化[J]. 中国药房, 2011, 0(7): 654-657 |
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| 作者姓名： | 李隆云 陈大霞 李泉森 |
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| 作者单位： | 重庆市中药研究院中药种植研究所;重庆市中药良种选育与评价工程技术研究中心; |
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| 基金项目： | 国家科技攻关计划课题(2001BA701A32;2004BA604A04); 重庆市科技计划项目(6455) |
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| 摘 要： | 目的:对仙茅随机扩增多态性DNA(RAPD)反应体系进行优化筛选。方法:以仙茅基因组DNA为模板,通过单因子试验就RAPD反应体系中主要参数(Mg2+、dNTP、引物、模板、TaqDNA聚合酶)对扩增结果的影响进行研究,并找出各自的最适条件。结果:建立了适合仙茅RAPD分析的反应体系,即25μL反应体系中各组分的最适浓度分别为:1×PCRbuffer、2.0mmol·L-1Mg2+、200μmol·L-1dNTP、40ng引物、20ng模板、1IUTaqDNA聚合酶。结论:利用此优化反应体系对仙茅进行RAPD-PCR扩增,可得到清晰的图谱,证实了该体系稳定、可靠。
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| 关 键 词： | 仙茅 随机扩增多态性DNA技术 反应体系 优化 |
Optimization of the RAPD Reaction System for Curculigo orchioides |
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| LI Long-yun,CHEN Da-xia,LI Quan-senLI Long-yun,LI Quan-sen. Optimization of the RAPD Reaction System for Curculigo orchioides[J]. China Pharmacy, 2011, 0(7): 654-657 |
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| Authors: | LI Long-yun CHEN Da-xia LI Quan-senLI Long-yun LI Quan-sen |
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| Affiliation: | LI Long-yun,CHEN Da-xia,LI Quan-sen(Institute of Material Medical Planting,Chongqing Academy of Chinese Materia Medica,Chongqing 400065,China)LI Long-yun,LI Quan-sen(Chongqing Engineering Research Center for Fine Variety Breeding Techniques of Chinese Materia Medica,China) |
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| Abstract: | OBJECTIVE： To study the optimization of RAPD reaction system for Curouligo orchioides. METHOD： Using genomic DNA extracted from C orchioides as template, the effects of the main parameters of RAPD reaction system （Mg2＋, dNTP, primer, template DNA, Taq DNA polymerase） on amplification results were tested by single factor experiment to determine its optimal levels. RESULTS： A suitable reaction system for C. orchioides was established, i.e. 25μL amplification reactions system containing 1×PCR buffer, 2.0 mmol·L^-1Mg2＋, 200μmol·L^-1 dNTP, 40 ng primer, 20 ng template DNA, 1 IU Taq DNA polymerase. CONCLUSION： This optimal reaction system can be used for RAPD-PCR amplification of C. orchioides. Obtained clear spectrum confirms the reliability of RAPD reaction system. |
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| Keywords: | Curculigo orchioides RAPD Reaction system Optimization |
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