观赏山梨叶片总RNA的提取及质量分析

为了从富含多糖和多酚等物质的观赏山梨幼叶中提取和分离出高质量的RNA,利用Trizol法、改良的Trizol法、异硫氰酸胍法、硼砂-CTAB法和硼砂-SDS法提取观赏山梨叶片总RNA。通过RNA产率、纯度、电泳图谱和RT-PCR结果等分析来确立适用于观赏山梨RNA分离的方法。结果表明,硼砂-SDS法提取的RNA呈现出28S rRNA,18S rRNA和5S rRNA三条较清晰的条带,RNA的得率和纯度均较高、很少有降解。RT-PCR结果进一步证实了硼砂-SDS法提取的观赏山梨叶片总RNA可用于分子生物学的下游试验。此方法简便易行,成本也较低,适合于富含多糖、多酚植物材料RNA的提取。另外4种方法获得的RNA得率、纯度较低,降解严重,有较多小分子和盐存在,因此不适合用于观赏山梨总RNA的提取。

Methods for RNA isolating from leaves of Pyrus ussuriensis Maxim

In order to isolate high quality RNA from young leaves of Pyrus ussuriensis Maxim,in which polyphenols and polysaccharide are rich,The authors used five methods as follows: Trizol reagent;improved Trizol reagent;Guanidine Thiocyanate;borax-CTAB;borax-SDS,and established the most suitable isolation method of Pyrus ussuriensis Maxim RNA through the rate of yield,purity,electrophoresis and RT-PCR.The results showed that there were three clear bands-28S rRNA,18S rRNA and 5S rRNA and little RNA degradation,the rates of yield and purity were all higher by the method of borax-SDS.Isolated RNA was intact and had been proved suitable for further molecular application by RT-PCR results.It is believed that the borax-SDS method is especially useful for the RNA extraction of plant materials which have plenty of polysaccharide and polyphenols.The RNA obtained from other four methods had low rate of yield and purity,decomposed seriously,contained much small molecule and salinity,so they are all not suitable for the RNA extraction from the leaves of Pyrus ussuriensis Maxim.