半夏叶片总RNA的提取研究

目的]为了获得高产高质的半夏RNA,为半夏分子生物学方面的研究打下基础。方法]采用改进的CTAB法提取半夏叶片RNA。用紫外分光光度计测定所提RNA样品的OD260、OD280和OD230的值,用RNA浓度计算公式:RNA(μg/μl)=OD260×40×稀释倍数/1000,求出RNA浓度。结果]紫外分光光度计的测定结果表明:OD260/OD280介于1.7～2.0之间。用改进的CTAB法提取的半夏叶片RNA质量较好,经变性胶电泳后,有较明显的3条带,分别是28S、18S和5S,28S、18S清晰可见,5S稍有弥散,但在亮度上28S与18S差别不大。结论]此方法简便易行,成本较低,适合于富含多糖、多酚植物材料的RNA提取。

Study on the Extraction of Total RNA from Pinellia ternata(Thunb.)Berit Leaves

Objective] The purpose was to obtain Pinellia ternata RNA with high yield and quality and provide a basis for the research on P. ternata in molecular biology. Method] The improved CTAB method was used to extract RNA from P. ternata leaves. The UV spectrophotometer was used to determine the values of OD260, OD280 and OD230 of the extracted RNA samples. The calculation formula of RNA concn., i.e. RNA (μg /μl)=OD260×40×diluting multiple/1 000 was used to calculate RNA concn. Result] The determination results of UV spectrophotometer showed that the ratio of OD260 to OD280 was in 1.7-2.0. The RNA extracted from P. ternata leaves by the improved CTAB method had better quality and had 3 more obvious bands after degenerative gel electrophiresis, which were 28S, 18S and 5S resp. The bands of 28S and 18S were clear and visible and that of 5S had a little diffusion, but the bands of 28S and 18S had little difference on brightness. Conclusion] The method was simple, convenient and easy to do and had low cost. It was suitable for extracing RNA from plant materials rich in polysaccharide and polyphenol.