一种改良的快速大量提取禾谷类作物成熟籽粒总RNA的方法

An Improved Method for Total RNA Extraction from Mature Seeds of Gramineous Grain Crops

禾谷类作物成熟籽粒富含多糖、蛋白质和脂类等成分,传统的提取方法所得的RNA大多不能满足下游操作的需求。此实验利用一种改良的CTAB法对小麦、水稻、玉米成熟籽粒总RNA进行提取,实验步骤简单。RNA得率高,完整性好,电泳条带清晰,OD260/OD280值在1.6～1.8之间。RT-PCR和Northern印迹分析显示该法所提RNA能够满足下游实验操作的要求。进一步分析发现,这种改良的CTAB法对玉米籽粒RNA的提取效果优于小麦和水稻,更适于玉米籽粒RNA的提取。

Conventional methods for RNA extraction are not feasible to mature seeds of gramineous crops for the enrichment of polysaccharide,protein and lipid.In this paper an improved method for isolating total RNA from the seeds of wheat,rice and maize is introduced.Comparing with the Conventional methods for RNA extraction,the improved method was of high purity （with the ratio of OD 260 /OD 280 1.6~1.8） and integrity and the results of RT-PCR and Northern blot showed that the RNA extracted by this improved method can meet the experimental requirements for high quality RNA,such as microarray,small RNA library construction for subsequent Illumina sequencing.