芹菜RAPD反应体系的优化研究
Chinese Agricultural Science Bulletin(2011)
Abstract
本试验以一般RAPD反应程序为基础,采用单因素递进筛选方法,针对TaqDNA聚合酶、Mg2+、dNTPs、随机引物和DNA模板5个主要影响因素,分别设置5个不同浓度梯度,对芹菜进行RAPD-PCR扩增,建立了芹菜RAPD技术最优体系。结果表明:25μL反应体系中含TaqDNA聚合酶1.0U、Mg2+3.0mM、dNTPs0.2mM、引物28ng、模板DNA70ng,10×PCRBuffer2.5μL;扩增程序为:94℃预变性5min,94℃变性1min,36℃退火1min,72℃延伸1min,进行42个循环,最后72℃延伸10min。
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Key words
reaction system,celery,optimization,RAPD
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