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 impuri...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.展开更多
文摘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.
