| 黄芪ISSR-PCR反应体系的建立及优化 |
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| 引用本文： | 张茹,刘亚令,梁建萍,张鹏飞.黄芪ISSR-PCR反应体系的建立及优化[J].山西农业科学,2014(1):9-11. |
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| 作者姓名： | 张茹 刘亚令 梁建萍 张鹏飞 |
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| 作者单位： | [1]山西农业大学生命科学学院,山西太谷030801 [2]山西农业大学园艺学院,山西太谷030801 |
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| 基金项目： | 山西省科技攻关项目（20110313002-2,20130311022-6）;山西农业大学科技创新基金项目（2010003） |
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| 摘 要： | 为获得最优黄芪ISSR-PCR反应体系,以山西省浑源县官儿乡蒙古黄芪为试验材料,利用单因素试验法对反应体系中的4个因素(模板DNA,dNTPs,Taq DNA聚合酶,ISSR引物)分别设置浓度梯度进行体系优化。结果表明,20μL黄芪ISSR-PCR最优体系为:模板DNA质量浓度6.0 ng/μL,dNTPs浓度0.03 mmol/L,Taq DNA聚合酶浓度0.05 U/μL,引物浓度0.80 mmol/L。最后用不同产地的黄芪检测该体系,结果发现,PCR产物多态性良好。优化体系的确立为今后黄芪种质资源的分子评价奠定了技术基础。
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| 关 键 词： | 黄芪 梯度 ISSR PCR |
Establishment and Optimization of ISSR-PCR Reaction System for Astragalus |
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| ZHANG Ru;LIU Ya-ling;LIANG Jian-ping;ZHANG Peng-fei.Establishment and Optimization of ISSR-PCR Reaction System for Astragalus[J].Journal of Shanxi Agricultural Sciences,2014(1):9-11. |
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| Authors: | ZHANG Ru;LIU Ya-ling;LIANG Jian-ping;ZHANG Peng-fei |
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| Institution: | ZHANG Ru;LIU Ya-ling;LIANG Jian-ping;ZHANG Peng-fei(College of Life Science, Shanxi Agricultural University, Taigu 030801, China;College of Horticulture, Shanxi Agricultural University, Taigu 030801, China) |
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| Abstract: | To get the optimization ISSR-PCR reaction system of Astragalus,the ISSR-PCR system through the single factor experiment was established and optimized.The results show that the best reaction system is a 20 μL volume ISSR-PCR system with DNA temphte of 6.0 ng/μ L,dNTPs of 0.03 mmol/L,TaqDNA polymerase of 0.05 U/μL,primer of 0.80 mmol/L and the annealing temperature of 49 ℃.Finally,concluded that this optimal system of ISSR-PCR was established successfully after tested using the Astragalus membranaceus from provinces of Hebei,Jilin,Heilongjiang,Shanxi,Inner-mongoulia,Shandong,Shaanxi,and Gansu. |
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| Keywords: | Astragalus gradient ISSR PCR |
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