[1]毛伟,李培英,孙宗玖.偃麦草SRAP-PCR反应体系优化[J].北方园艺,2015,39(02):91-97.[doi:10.11937/bfyy.201502026]
　MAO Wei,LI Pei-ying,SUN Zong-jiu.Optimization of SRAP-PCR System on Elytrigia repens[J].Northern Horticulture,2015,39(02):91-97.[doi:10.11937/bfyy.201502026]

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偃麦草SRAP-PCR反应体系优化

《北方园艺》[ISSN:1001-0009/CN:23-1247/S] 卷: 第39卷期数: 2015年02 页码: 91-97 栏目: 出版日期: 2015-01-30

Title:: Optimization of SRAP-PCR System on Elytrigia repens

作者:: 毛伟; 李培英; 孙宗玖; (新疆农业大学草业与环境科学学院，新疆草地资源与生态自治区重点实验室，新疆乌鲁木齐 830052)

Author(s):: MAO Wei; LI Pei-ying; SUN Zong-jiu; (College of Pratacultural and Environmental Science，Xinjiang Agricultural University，Key Laboratory of Grassland Resource and Ecology of Xinjiang，Urumqi，Xinjiang 830052)

关键词:: 偃麦草; SRAP; 体系优化

Keywords:: Elytrigia repens; SRAP; system optimization

DOI:: 10.11937/bfyy.201502026

文献标志码:: A

摘要:: 以偃麦草叶片DNA为模板，利用单因子和L₁₆(4⁵)正交实验设计对影响偃麦草SRAP-PCR反应效果的Mg²⁺、引物、dNTPs、DNA、Taq DNA聚合酶5种因素进行优化，并比较了不同退火温度对扩增反应的影响，通过综合比较分析建立偃麦草SRAP-PCR的优化反应体系。结果表明：优化的偃麦草SRAP-PCR总体系20μL中，Mg²⁺1.75 mmol/L，引物 0.15 μmol/L，dNTPs 0.20 mmol/L，DNA 50 ng，Taq DNA聚合酶 0.75 U，2 μL 10 ×PCR buffer；Mg2+和引物浓度对扩增效果影响最大，DNA浓度影响最小；采用该体系对32份偃麦草进行验证，扩增结果清晰稳定，此体系的建立为利用SRAP分子标记进行偃麦草遗传多样性、抗性标记等研究奠定了技术基础

Abstract:: With genetic DNA of the Elytrigia repens as template，single factor and orthogonal design were applied to optimize the concentration of Mg²⁺,primer,dNTPs,DNA，Taq DNA polymerase for SRAP-PCR system of Elytrigia repens，the effect of different annealing temperature on reaction was studied by comprehensive analysis，the optimal system of SRAP-PCR was established.The results showed that the optimal system was Mg²⁺ 1.75 mmol/L，primer 0.15 μmol/L，dNTPs 0.20 mmol/L,DNA 50 ng/20μL，Taq DNA polymerase 0.75 U，2 μL 10 ×PCR in 20μL，concentrations of Mg²⁺ and primer had the highest effect on reaction，DNA concentrations had the least effect.The test of optimal system on 32 kinds of Elytrigia repens were done，the result was steady and clear，the establishing of optimal system made the base for the studies including genetic diversity and resistance marker and so on.

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备注/Memo

第一作者简介：毛伟（1986-），女，重庆人，硕士研究生，研究方向为草坪及草种资源评价及育种。E-mail：569646469@qq.com.责任作者：李培英（1975-），女，内蒙古乌兰察布市察右前旗人，博士，副教授，现主要从事草坪及草种资源评价和遗传育种等科研与教学工作。E-mail：nmlpy_1234@sina.com.基金项目：新疆自然科学基金资助项目(2012211A060)。

更新日期/Last Update: 2015-02-24