| 牛筋草ISSR-PCR反应体系的建立与优化 |
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| 引用本文： | 张杨,张朝贤,王金信,魏守辉,黄红娟.牛筋草ISSR-PCR反应体系的建立与优化[J].植物保护,2012,38(3):90-94. |
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| 作者姓名： | 张杨 张朝贤 王金信 魏守辉 黄红娟 |
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| 作者单位： | 1. 山东农业大学植物保护学院,泰安,271018
2. 中国农业科学院植物保护研究所,中国农业科学院杂草鼠害生物学与治理重点开放实验室,北京100193 |
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| 基金项目： | 转基因生物新品种培育重大专项(2009ZX08012-025B) |
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| 摘 要： | 以牛筋草地上组织为材料,采用正交优化和单因子试验两种方法对影响牛筋草ISSR-PCR体系的Mg2+浓度、dNTP浓度、Taq DNA聚合酶浓度、引物浓度和DNA模板浓度5个因素进行优化试验,建立适合牛筋草ISSR-PCR的反应体系.结果表明,牛筋草ISSR-PCR反应体系的最佳条件为:Mg2+1.75 mmol/L,dNTP 0.4 mmol/L,Taq DNA聚合酶1 U/25 μL,引物0.3μmol/L,模板DNA 80 ng/25μL,10×PCR Buffer 2.5 μL/25 μL.利用优化体系进行牛筋草的ISSR-PCR反应,可获得稳定性高、重复性好、背景清晰的电泳结果.
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| 关 键 词： | 牛筋草 ISSR-PCR 正交设计 单因子试验 |
Establishment and optimization of ISSR-PCR reaction system for Eleusine indica |
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| Zhang Yang,Zhang Chaoxian,Wang Jinxin,Wei Shouhui,Huang Hongjuan.Establishment and optimization of ISSR-PCR reaction system for Eleusine indica[J].Plant Protection,2012,38(3):90-94. |
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| Authors: | Zhang Yang Zhang Chaoxian Wang Jinxin Wei Shouhui Huang Hongjuan |
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| Institution: | 1.College of Plant Protection,Shandong Agricultural University,Tai’an 271018,China; 2.Institute of Plant Protection,Key Laboratory of Weed and Rodent Biology and Management, Chinese Academy of Agricultural Sciences,Beijing 100193,China) |
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| Abstract: | The suitable ISSR-PCR reaction system of Eleusine indica was established by optimizing the concentrations of Mg2+,dNTP,Taq DNA polymerase,primers and template DNA in the ISSR-PCR reaction system with orthogonal test and single-factor test.The best conditions were as followed: 1.75 mmol/L Mg2+,0.4 mmol/L dNTP,1 U/25 μL Taq DNA polymerase,0.3 μmol/L primer,80 ng/25 μL of DNA template and 2.5 μL/25 μL of 10×PCR Buffer.With the optimized ISSR-PCR reaction system,the good results with high stability,reproducibility and clear background of electrophoresis could be obtained. |
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| Keywords: | Eleusine indica ISSR-PCR orthogonal design single factor test |
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