| 3种实蝇基因组DNA提取及ISSR-PCR反应体系的建立 |
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| 引用本文： | 余德亿,姚锦爱,胡进锋,黄鹏,方大琳,陈峰.3种实蝇基因组DNA提取及ISSR-PCR反应体系的建立[J].农学学报,2011(5). |
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| 作者姓名： | 余德亿 姚锦爱 胡进锋 黄鹏 方大琳 陈峰 |
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| 作者单位： | 福建省农业科学院植物保护研究所;福建省农作物害虫天敌资源工程技术研究中心; |
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| 基金项目： | 福建省科技重大专项专题“花卉产业化配套技术研发-盆栽花卉病虫害防治技术研究”(2010NZ0003-2-3); 福建省属公益类科研院所基本科研专项计划项目“漳州出口盆栽花卉病虫害种类调查和防治新技术研究”(2010R1026-4); 福建省科技创新平台建设项目“福建省农作物害虫天敌资源工程技术研究中心”(2008N2002) |
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| 摘 要： | 为探讨高品质实蝇基因组DNA提取,研究模板DNA浓度、引物浓度、TaqDNA聚合酶用量、dNTP浓度、退火温度及时间对ISSR-PCR扩增结果的影响,以3种实蝇为材料,建立通用且稳定的实蝇ISSR-PCR反应体系。结果表明:获得了高品质实蝇基因组DNA;确立了通用且稳定的实蝇ISSR-PCR反应体系:10×PCRBuffer2.5μL,模板DNA50ng,引物0.25μmol/L,TaqDNA聚合酶0.50U,dNTP200μmol/L,最后加ddH2O至25μL;明确了ISSR-PCR扩增程序:94℃预变性5min,94℃变性30s,52.4℃退火45s,72℃延伸90s,循环36次,最后72℃延伸7min,4℃保存。体系的建立弥补了实蝇传统形态检测的不足,为快速准确鉴定、种群异质性及遗传多样性分析奠定了基础。
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| 关 键 词： | 实蝇 基因组DNA提取 ISSR-PCR反应体系 建立 |
Genomic DNA Extraction of Three Kinds of Fruit Flies and Their ISSR-PCR Reaction System Establishment |
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| Yu Deyi ,Yao Jin ' ai ,Hu Jinfeng ,Huang Peng ,Fang Dalin ,Chen Feng.Genomic DNA Extraction of Three Kinds of Fruit Flies and Their ISSR-PCR Reaction System Establishment[J].Journal of Agriculture,2011(5). |
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| Authors: | Yu Deyi Yao Jin ' ai Hu Jinfeng Huang Peng Fang Dalin Chen Feng |
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| Institution: | Yu Deyi 1,2,Yao Jin ' ai 1,Hu Jinfeng 1,Huang Peng 2,Fang Dalin 2,Chen Feng 1 ( 1 Institute of Plant Protection,Fujian Academy of Agricultural Sciences,Fuzhou 350013,Fujian,China,2 The Engineering Technology Research Center of Fujian Crops Pests Natural Enemies Resources,China) |
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| Abstract: | In order to explore the extraction of high-quality genomic DNA of three kinds of fruit flies, and to study the effect of template DNA concentration, primer concentration, the amount of Taq DNA polymerase, dNTP concentration, annealing temperature and annealing time on ISSR-PCR (inter-simple sequence repeat-polymerase chain reaction) amplified results, the fruit flies ' general and stable ISSR-PCR reaction system was established. It was found that the fruit flies ' genomic DNA with high quality was obtained,... |
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| Keywords: | Fruit Flies the Extraction of Genomic DNA ISSR-PCR Reaction System Establishment |
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