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Position: Home > Articles > Total DNA Extraction from Artemisia argyi and Optimization on ISSR-PCR Reaction System Guizhou Agricultural Sciences 2015 (6) 165-170

中药艾总DNA的提取及ISSR-PCR反应体系的优化

作  者:
王惠君;王文泉;卢诚;王海燕;陈新
单  位:
中国热带农业科学院;琼州学院
关键词:
艾;分子标记;ISSR;传统中药
摘  要:
为探索艾的遗传多样性,采用正交试验方法 L16(45)与单因素分析法相结合对其PCR反应的5个主要因素(Mg2+、引物、模板DNA、dNTPs以及Taq酶)在4个水平上分别进行优化。结果表明:得到最佳的反应体系(20μL)为模板DNA 60ng,Mg2+2.5mmol/L,引物0.8μmol/L,dNTPs 0.15mmol/L,Taq酶1.0U。通过统计数据分析,对艾ISSR-PCR反应的影响程度依次为Mg2+浓度>引物>模板DNA>dNTPs浓度>Taq酶。
译  名:
Total DNA Extraction from Artemisia argyi and Optimization on ISSR-PCR Reaction System
作  者:
WANG Huijun;WANG Wenquan;LU Cheng;WANG Haiyan;CHEN Xin;Qiongzhou University;Chinese Academy of Tropical Agricultural Sciences/Institute of Tropical Bioscience and Biotechnology;
关键词:
Artemisia argyi;;molecular marker;;ISSR;;traditional Chinese medicine
摘  要:
In order to explore the genetic diversity of A.argyi,an orthogonal experimental design was employed combined with single factor analysis to optimize the five main factors(Mg2+,primers,template DNA,dNTPs and Taq DNA enzyme)of PCR reaction in four levels.Results:The optimal ISSR-PCR reaction system:20μL system,template DNA 60 ng,Mg2+2.5 mmol/L,primer 0.8μmol/L,dNTPs0.15mmol/L,Taq DNA enzyme 1.0 U.The results of range analysis showed that the order of the influence degree to ISSR was Mg2+> primer>template DNA> dNTPs> Taq DNA enzyme.

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