| 实蝇RAPD反应体系的构建与优化 |
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| 引用本文： | 张长禹,孟建玉,张小亚,雷朝亮.实蝇RAPD反应体系的构建与优化[J].中国农学通报,2007,23(1):58-058. |
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| 作者姓名： | 张长禹 孟建玉 张小亚 雷朝亮 |
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| 作者单位： | 华中农业大学昆虫资源研究所,武汉,430070 |
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| 摘 要： | 以几种常见的实蝇为材料，研究了实蝇RAPD分析过程中的影响因素，包括模板DNA浓度、dNTP、引物、Taq酶、变性时间、循环次数、退火温度等，建立了适于实蝇RAPD分析的PCR反应体系：在25μl反应体系中，模板DNA用量为80ng，MgCl 2浓度为2.0mM；四种dNTP浓度各为0.2 mM；Taq引物浓度为0.5μM；酶用量为1U；扩增程序：先94℃预变性3min，再94℃变性45s、36℃退火1min、72℃延伸2min共40个循环，最后延伸10min。
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| 关 键 词： | 绞股蓝 绞股蓝 叶龄 皂甙 |
| 修稿时间： | 2006-10-202006-10-30 |
Optimization For RAPD Reaction System in Fruit Flies (Diptera:Tephritidae) |
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| Zhang ChangYu,Meng JianYu,Zhang XiaoY,Lei Chaoliang.Optimization For RAPD Reaction System in Fruit Flies (Diptera:Tephritidae)[J].Chinese Agricultural Science Bulletin,2007,23(1):58-058. |
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| Authors: | Zhang ChangYu Meng JianYu Zhang XiaoY Lei Chaoliang |
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| Institution: | Institute of Insect Resources, Huazhong Agricuhural University, Wuhan 430070 |
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| Abstract: | The factors influencing RAPD Analysis, including the concentration of DNA template, Mg2+, dNTP, primers, Taq polymerase, denaturing time, Thermal cycles and annealing temperature in fruit flies were studied. An optimal PCR system for RAPD in fruit flies has been found: in 25μl reaction solution, contained 80ng DNA template, 2.0mMMg2+, 0.2 mMdNTP, 0.5μMprimers, 1U Taq polymerase.The amplification program was devised: 94℃ for 3min; denaturing at 94℃ for 45sec, primer annealing at 36℃ for 1min, extension at 72℃ for 2min, 40cyclesm, at last extension at 72℃ for 10min. |
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| Keywords: | RAPD |
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