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作 者:樊洪泓[1] 李廷春[1] 林毅[1] 蔡永萍[1]
机构地区:[1]安徽农业大学生命科学学院,安徽合肥230036
出 处:《激光生物学报》2007年第1期105-108,共4页Acta Laser Biology Sinica
基 金:安徽省教育厅自然科学重点科研项目(2006KJ054A)
摘 要:采用TRIZOL法、异硫氰酸胍法、Tris-硼酸法和改良的RNA提取方法提取石斛的总RNA,并通过凝胶电泳、紫外分光光度法检测提取的RNA样品的品质。研究结果表明:改良的RNA提取方法提取的RNA具有28S rRNA和18S rRNA两条清晰的条带,且无降解。OD260nm/OD280nm接近2.0,具有较高的纯度。其它三种方法获得的RNA品质较差,有降解和弥散现象。将改良的RNA提取方法提取的RNA逆转录成cDNA,经RAPD扩增,出现清晰的条带,进一步证明改良的RNA提取方法提取的RNA具有很高的纯度,可以满足进一步分子生物学研究的要求。Total RNA was isolated from Dendrobium displaying different status by using the methods of TRIZOL Regent, Guanidine Thiocyanate, Tris-H3 BO4 , and improved RNA isolation. The quality of total RNA was analyzed through gel electrophoresis and UV spectrometer. The results indicated that the RNA isolated by the improved RNA isolation method showed clear bands of 28 S rRNA and 18 S rRNA, and the value of OD260nm/OD280nm was close to 2.0. RNA isolated by the other three methods degraded and dispersed in some degrees. RNA isolated by the improved RNA isolation method could be reverse to cDNA. The cDNA was used for RAPD amplifying and two clear bands could be observed in agarose gel. These results demonstrate that the quality and purity of the RNA obtained by the improved RNA isolation method can meet the demands of molecular biology experiment.
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