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中国农学通报

中国农学通报 ›› 2006, Vol. 22 ›› Issue (6): 35-35.

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苎麻RAPD反应体系的构建与优化

刘立军,彭定祥,蒙祖庆   

  • 出版日期:2006-06-05 发布日期:2006-06-05

Optimization for RAPD Reaction System in Ramie(Boehmeria nivea L.Guad)

Liu Lijun, Peng Dingxiang, Meng Zuqing   

  • Online:2006-06-05 Published:2006-06-05

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摘要： 以苎麻品种为材料，研究了苎麻RAPD分析过程中的影响因素，包括10×PCR Buffer、模板浓度、Mg2+、dNTP、引物、Taq酶、循环次数、退火温度等，建立了适于苎麻RAPD分析的PCR反应体系：即在25μl反应体系中，引物浓度为0.4uM；模板DNA的用量为60~100ng；Mg2+浓度为1.8mM；dNTP浓度为0.2 mM；Taq酶用量为1U。适宜的扩增程序为先94℃变性4min，再94℃变性45s、38℃复性45s、72℃延伸2min共36个循环，最后72℃延伸5min，4℃保存。

关键词: 高油玉米, 高油玉米, 高产优质栽培, 数学模型, 栽培模式

Abstract: The factors influencing RAPD Analysis, including 10×PCR Buffer, the concentration of DNA template, Mg2+, dNTP, primers, Taq polymerase, Thermal cycles and annealing temperature in Ramie were studied. PCR system for RAPD in Ramie has been found: in 25μl reaction solution, contained 0.4μM random primers, 60~100ng DNA template, 1.8mM Mg2+, 0.2 mM dNTP, 1U Taq polymerase. The suitable procedure is one cycle 94℃ for 4min; denaturing at 94℃ for 45s; primer annealing at 38℃ for 45s, extension at 72℃ for 2min, 36 cycles; extension at 72℃ for 5min at last and then remain at 4℃.