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| 本氏针茅ISSR-PCR反应体系的建立及引物筛选 | |
| 引用本文： | 俞靓,井赵斌,魏琳,程积民.本氏针茅ISSR-PCR反应体系的建立及引物筛选[J].中国草地学报,2012,34(1):77-83. |
| 作者姓名： | 俞靓 井赵斌 魏琳 程积民 |
| 作者单位： | 1. 西北农林科技大学资源与环境学院,陕西杨凌,712100 2. 西北农林科技大学动物科技学院,陕西杨凌,712100 3. 中国科学院水利部水土保持研究所,陕西杨凌,712100 |
| 基金项目： | 国家现代农业产业技术体系建设-咸阳牧草试验站;中国科学院院地合作专项宁夏南部山区生态保育型集约化养殖技术模式转化与示范资金资助 |
| 摘 要： | 以CTAB法提取的本氏针茅(Stipa bungeana Trin.)基因组DNA为模板,采用正交试验设计和单因素分析相结合的方法,对影响本氏针茅ISSR-PCR反应体系中的DNA、Taq酶、dNTPs、引物和Mg2+进行优化,旨在建立适合本氏针茅ISSR-PCR分析的最佳反应体系。结果表明,在20μL的反应体系中各组分的浓度分别为:DNA(20ng/μL)2.5μL、Taq DNA酶(5U/μL)0.1μL、dNTPs(2.5mmol/L)1.6μL、引物(10μmol/L)2.3μL、Mg2+(25mmol/L)1.4μL、10×Buffer 2.5μL、ddH2O 9.6μL。经过体系验证和引物筛选试验表明该体系适于本氏针茅遗传多样性分析,该体系的建立为本氏针茅种质资源遗传多样性研究提供了理论基础。 |
| 关 键 词： | 本氏针茅 ISSR 体系优化 |
Establishment and Optimization of the ISSR-PCR Reaction System in Stipa bungeana Trin. |
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| YU Jing , JING Zhao-bin , WEI Lin , CHENG Ji-min.Establishment and Optimization of the ISSR-PCR Reaction System in Stipa bungeana Trin.[J].Chinese Journal of Grassland,2012,34(1):77-83. | |
| Authors: | YU Jing JING Zhao-bin WEI Lin CHENG Ji-min |
| Institution: | 1.College of Resources and Environment,Northwest A &F University,Yangling 712100,China; 2.College of Animal Science and Technology,Northwest A &F University,Yangling 712100, China;3.Institute of Soil and Water Conservation,Chinese Academy of Sciences and Ministry of Water Resources,Yangling 712100,China) |
| Abstract: | An improved CTAB method was used to extract the genomic DNA of Stipa bungeana,an orthogonal design was combined with a factor analysis method to optimize the ISSR-PCR reaction system of S.bungeana.Five main factors including DNA,Taq Polymerase,dNTPs,Mg2+ and primers were optimized in order to establish a suitable PCR reaction system for Stipa bishbungeana.The results indicated that the concentration of each components in total 20μl reaction system were: DNA(20ng/μL)2.5μL,Taq polymerase(5U/μL) 0.1μL,dNTPs(2.5mmol/L) 1.6μL,primer(10μmol/L)2.3μL,Mg2+(25mmol/L) 1.4μL,10×Buffer 2.5μL,ddH2O 9.6μL.This reaction system is suitable for the genetic diversity analysis of Stipa bungeana.To establish the SRAP-PCR reaction system provides a theory basis for the genetic diversity analysis of S.bungeana germplasm resources. |
| Keywords: | Stipa bungeana Trin ISSR System optimization |
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