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×PCRBuffer、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0.2mM;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。
The factors influencing RAPD Analysis, including 10× PCR Butter, the concentration of DNA template, Mg^2＋, 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 Mg^2＋, 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℃.
出处
《中国农学通报》
CSCD
2006年第6期35-39,共5页
Chinese Agricultural Science Bulletin
基金
国家863项目麻类作物高效育种技术及优质高产多抗新品种选育(2001AA241121)。
