百脉根总RNA提取方法的优化

目的]为百脉根的分子生物学研究提供基础资料。方法]以百脉根叶片为材料,比较异硫氰酸胍法、SDS-LiCl法及SDS-LiCl改进法提取总RNA的效果并探讨活性炭、抗坏血酸对RNA提取质量的影响。结果]异硫氰酸胍法所提取的百脉根总RNA降解严重,盐类去除不充分,该方法不适合百脉根总RNA的提取;SDS-LiCl法提取的总RNA有一定程度降解,加入液体抗坏血酸会降低百脉根RNA的提取质量;SDS-LiCl改进法提取的总RNA较为完整,有较高的纯度,在研磨过程中加入固体抗坏血酸能够提高百脉根总RNA的提取质量。活性炭对百脉根总RNA的提取无影响。结论]研磨时加入固体抗坏血酸的SDS-LiCl改进法能提取出高质量的百脉根总RNA。

Optimization Method of Total RNA Extraction from Lotus japonicus

Objective] The aim was to provide the basic data for molecular biology research of Lotus japonicus.Method] With the L.japonicus leaves as material, the effect of total RNA extraction using different methods of guanidine thiocyanate method,SDS-LiCl method and SDS-LiCl improved method was compared and the influence of the active carbon and ascorbic acid on the quality of RNA extraction was discussed.Result] The total RNA extracted by guanidine thiocyanate method from L.japonicus was degraded seriously,the salt removal was not sufficient and the method was not suitable for the total RNA extraction from L.japonicus;the total RNA extracted by SDS-LiCl method was degraded to some extent and adding liquid ascorbic acid could decrease the extraction quality of RNA from L.japonicus;the total RNA extracted by SDS-LiCl improved method was comparatively complete and had higher purity,and adding solid ascorbic acid when grinding could improve the extraction quality of the total RNA from L.japonicus.Conclusion] The SDS-LiCl improved method of adding solid ascorbic acid when grinding could extract the total RNA with high quality from L.japonicus.