用改良CsCl密度梯度离心法提取石蒜叶片和鳞茎总RNA  

Total RNA extracted from leaf and bulb of Lycoris radiata by an improved method of CsCl density gradient centrifugation

作　　者：江玉梅[1] 汪仁[1] 束晓春[1] 佟金凤[1] 彭峰[1] 夏冰[1] 

机构地区：[1]江苏省.中国科学院植物研究所(南京中山植物园),江苏南京210014

出　　处：《植物资源与环境学报》2008年第3期78-80,共3页Journal of Plant Resources and Environment

基　　金：国家自然科学基金资助项目(30700057);江苏省植物迁地保护重点实验室开放基金项目(KF07001)

摘　　要：The total RNA is extracted from leaf and bulb of Lycoris radiata(L’Hér.)Herb.by an improved method of CsCl density gradient centrifugation.The suitable amount of initial sample is 1.0 g.In order to remove impurities and purify the total RNA,the crude extract of RNA dissolved in lysis buffer is extracted with the mixed solution of chloroform and isoamyl alcohol(V ∶V,24 ∶1),then the total RNA is precipitated.The purity,integrity and quality of the total RNA are detected by ultraviolet spectrophotometer,agarose gel electrophoresis and RT-PCR method,respectively.The results show that polysaccharide can be removed effectively from the total RNA extract by the improved method of CsCl density gradient centrifugation.The purity andintegrityof the total RNA of L.radiata are better,and the ratio of A260/A280 is 1.85-2.00.It is concluded that the total RNA of L.radiata bulb extracted by the improved method can satisfy the demand of molecular biology experiments.The total RNA is extracted from leaf and bulb of Lycoris radiata （ L＇ Her. ） Herb. by an improved method of CsCl density gradient centrifugation. The suitable amount of initial sample is 1.0 g. In order to remove impurities and purify the total RNA, the crude extract of RNA dissolved in lysis buffer is extracted with the mixed solution of chloroform and isoamyl alcohol （ V ： V, 24 ： 1 ）, then the total RNA is precipitated. The purity, integrity and quality of the total RNA are detected by ultraviolet spectrophotometer, agarose gel electrophoresis and RT-PCR method, respectively. The results show that polysaccharide can be removed effectively from the total RNA extract by the improved method of CsC1 density gradient centrifugation. The purity and integrity of the total RNA of L. radiata are better, and the ratio of A260/A280 is 1.85 - 2.00. It is concluded that the total RNA of L. radiata bulb extracted by the improved method can satisfy the demand of molecular biology experiments.

关 键 词：石蒜 RNA CsCl密度梯度离心法 RT—PCR 

分 类 号：Q946-33[生物学—植物学] Q522