麦冬根中总RNA的快速提取

Quick Extraction of Total RNA from the Root of Ophiopogon japonicus (L.F.) Ker-Gawl

目的:从富含多糖、多酚的麦冬根部组织中快速提取总RNA。方法:采用改进的苯酚法,提取液的配制为5%SDS、1mol/LNaAc(pH4.1)、20%HAC、0.1%PVP。结果:采用该方法提取的麦冬总RNA纯度高、完整性好,电泳条带清晰。通过琼脂糖凝胶电泳与紫外吸光度测定产量与纯度,麦冬根部组织总RNA的吸光值D260nm/D280nm值大于1.8,D260nm/D230nm值大于2.0,麦冬块根与不定根RNA的平均产量分别为79.716和76.144μg/g(鲜重)。结论:用本方法提取的RNA可用于后继的抑制消减杂交试验。

Objective：To develop an effective method for total RNA isolation from the root tissue of Ophiopogon japonicus（L.F.） Ker-Gawl rich in polysaccharides and polyphenolics.Methods：Improved phenol method was applied.RNA extraction buffer was composed of 5% SDS,1 mol /L NaAc（pH4.1）,20% HAC,and 0.1% PVP.Results：The RNA extraction protocol presented might be used to efficiently isolate high quality RNA with high purity and integrity,clear electrophoresis.Yield and purity were monitored by gel electrophoresis and by UV absorbance.D260nm/D280nm and D260nm/D230nm absorbance ratio of root tissue RNA of O.japonicus were higher than 1.8 and 2.0 respectively.Average yields of RNA of touberous root and adventitious root were about 79.716 and 76.144 μg /g（FW） respectively.Conclusion：The RNA extracted by this method was of sufficient quality for use in SSH reaction.