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作 者:刘胜洪[1] 刘文[1] 刘明峰[1] 杨凤婷[1] 刘庆生[1] 梁红[1]
机构地区:[1]仲恺农业工程学院生命科学学院,广州510225
出 处:《生物技术通报》2011年第9期171-175,共5页Biotechnology Bulletin
基 金:广东省自然科学基金项目(8151022501000010)
摘 要:在总核酸提取方法(PS法)的基础上,经多次实践改进,得出一种以高盐低pH的HAc-NaAc缓冲体系提取总核酸的简便方法,可以从富含多糖、多酚的猕猴桃叶片和花蕾中提取同时含有DNA和RNA的总核酸。所得的总核酸在LiCl溶液中选择性沉淀RNA,从而有效地分离出DNA和RNA样品。紫外分光光度法和琼脂糖凝胶电泳分析表明,所提取的DNA和RNA具有较高的纯度和完整性。通过样品DNA的PCR和样品RNA的RT-PCR,认为所提取的DNA样品和RNA样品能够满足分子生物学试验的基本要求。Total nucleic acid was extracted through modified PS method from kiwifruit leaves and flower buds and the DNA and RNA were isolated from the nucleic acid sample.The experimental results showed that the kiwifruit leaves and flower buds,rich in polysaccharides and polyphenols,could be easily obtained nucleic acid by extraction solution with HAc-NaAc buffer system at low pH.Then the genomic DNA and total RNA were separated by selectively precipitating RNA with LiCl solution.Observations based on ultraviolet spectrophotometry and agarose gel electrophoresis showed high purity and integrity of the DNA and RNA.The DNA and RNA samples could meet the needs for PCR and RT-PCR analysis.
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