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Establishment and Optimization of ISSR-PCR Reaction System of Ligustrum lucidum Ait.
Author(s): LI Chuan-dai1, 2, CHEN Fei1, 2, WANG Tian-di1, 2, LIU Guo-min1, 2
Pages: 99-
104
Year: 2010
Issue:
2
Journal: Journal of Tropical Organisms
Keyword: GetLinkList(KeywordFilter('女贞; ISSR; 影响因子; 体系优化'); 'kw'; 'CJFQ');
Abstract: 利用单因子试验,测试了女贞ISSR-PCR反应体系中的主要影响因子。经过优化实验,建立了一套适合于女贞的稳定的ISSR-PCR反应体系:10×buffer2.5μL,2.0mmol·L-1的MgCl2,200μmol·L-1的dNTPs,1.5U的Taq酶,0.4的μmol·L-1的引物,10ng的DNA模板。PCR扩增程序为:94℃预变性4min,94℃变性40s,52~62℃退火45s,72℃延伸120s,进行35个循环,72℃延伸8min,4℃保存。应用该优化的ISSR-PCR实验体系对12份女贞种质材料进行了扩增,均能扩增出丰富稳定的条带。
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