吴茱萸ISSR-PCR反应体系优化 |
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引用本文: | 吴波,高丹,潘超美,张寿文.吴茱萸ISSR-PCR反应体系优化[J].安徽农业科学,2011,39(32):19686-19688. |
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作者姓名: | 吴波 高丹 潘超美 张寿文 |
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作者单位: | 1. 广州中医药大学中药学院,广东广州510006;江西中医学院江西省中药种质资源工程技术研究中心,江西南昌330004 2. 江西中医学院药学院,江西南昌,330004 3. 广州中医药大学中药学院,广东广州,510006 |
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基金项目: | 国家“十二五”科技支撑计划项目(2011BAI04B04) |
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摘 要: | 目的]建立吴茱萸ISSR-PCR最佳反应体系。方法]采用改良的CTAB法提取吴茱萸基因组DNA,研究模板DNA、dNTPs、Mg^2+、引物(U834)、Taq DNA聚合酶的浓度对ISSR-PCR扩增结果的影响。结果]20μl反应体系中含2μl 10×PCR buffer,0.15 mmol/LdNTPs,1.6 mmol/L Mg^2+,0.4 mmol/L引物,20 ng模板DNA,1.2 U Taq DNA聚合酶。结论]为研究吴茱萸种质资源遗传多样性和分子鉴定奠定了技术基础。
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关 键 词: | 吴茱萸 ISSR 反应体系 优化 |
Optimization for ISSR-PCR Reaction System in Evodia rutaecarpa |
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Institution: | WU Bo et al(College of Pharmacy,Guangzhou University of Chinese Medicine,Guangzhou,Guangdong 510006) |
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Abstract: | Objective]The optimal ISSR-PCR reaction conditions for Evodia rutaecarpa were determined.Method]The genomic DNA of Evodia rutaecarpa was extracted with improved CTAB method.The effects of concentration of template DNA,dNTPs,Mg2+,primer(U834) and Taq DNA polymerase were studied.Result]The optimum ISSR-PCR reaction system with total 20 μl reaction solution contained 2 μl 10×PCR buffer,0.15 mmol/L dNTPs,1.6 mmol/L Mg2+,0.4 mmol/L primer,20 ng template DNA and 1.2 U Taq DNA polymerase.Conclusion]The study laid the foundation for genetic diversity and molecular identification of germplasm resource in Evodia rutaecarpa. |
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Keywords: | Evodia rutaecarpa ISSR Reaction system Optimization |
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