山杏RAPD反应体系条件的优化

摘要/Abstract

摘要： 以山杏新鲜叶片为材料，研究了山杏RAPD分析过程中的影响因素，包括Taq酶、Mg2+、dNTP、引物、模板DNA浓度、变性时间、循环次数等，建立了适合山杏RAPD反应的PCR体系，即20μL反应体系中含有Taq酶1.0U、Mg2+ 2.0mM，四种dNTP各0.1mM，引物0.5μM，模板DNA50ng。扩增程序为：94℃预变性4mins；94℃变性1min，36℃退火1min，72℃延伸1.5mins，10个循环；94℃变性30s，36℃退火40s，72℃延伸60s，30个循环；最后72℃延伸5min

关键词: 土壤肥力, 土壤肥力, 烤烟, 硝酸盐, 积累

Abstract: The factors influencing RAPD analysis, including Taq polymerase, Mg2+, dNTP, primers, template DNA’s concentration, denaturing time and thermal cycles in Armeniaca. sibirica (L.) Lam. were studied. An optimal PCR system for RAPD in fresh leaves of A. sibirica (L.) Lam. has been found： in 20μL reaction solution, contained 1U Taq polymerase, 2.0mMMg2+, 0.1mM dNTP, 0.5μM primers, 50ng template DNA. The amplification program was devised： initial denaturing at 94℃ for 4mins; denaturing at 94℃ for 1mins, annealing at 36℃ for 1min, extension at 72℃ for 1.5mins, 10cycles; denaturing at 94℃ for 30s, annealing at 36℃ for 40s, elongation at 72℃ for 60s, 30 cycles; final extension at 72℃ for 5mins.

中图分类号:

S662.2

李振江,葛会波,,张学英,王勇. 山杏RAPD反应体系条件的优化[J]. 中国农学通报, 2007, 23(6): 169-169.

Li Zhenjiang, Ge Huibo,, Zhang Xueying, Wang Yong. Optimization for RAPD Reaction System of Armeniaca sibirica[J]. Chinese Agricultural Science Bulletin, 2007, 23(6): 169-169.

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