草莓果实总RNA提取方法的比较  

Comparison of Extraction Methods of Total RNA from Strawberry Fruits

作　　者：张卿[1] 刘帅[1] 邢宇[1] 曹庆芹[1] 秦岭[1] 

机构地区：[1]北京农学院农业应用新技术北京市重点实验室,北京102206

出　　处：《中国农学通报》2015年第31期146-149,共4页Chinese Agricultural Science Bulletin

基　　金：北京农学院青年基金重点项目"VIGS技术体系在果树上的建立"(QN2012001)

摘　　要：为建立草莓果实总RNA的提取方法,针对草莓成熟果实中富含多糖、多酚和色素等次级代谢物质,总RNA提取难度大的特点,比较改良的EASYspin植物RNA提取试剂盒法和改良的CTAB法提取草莓果实中总RNA的质量。通过琼脂糖凝胶电泳和核酸蛋白测定仪(Nano Drop 2000)检测2种方法提取总RNA的浓度、纯度及完整性等。改良的EASYspin植物RNA提取试剂盒法和改良的CTAB法都能够完成草莓果实总RNA的提取,电泳检测在28S和18S处呈2条清晰的条带,OD260/OD280值和OD260/OD230值均在2.0左右;改良的EASYspin植物RNA提取试剂盒法成本较高,且提取总RNA质量劣于改良的CTAB法,但是EASYspin植物RNA提取试剂盒法操作简便,安全可靠,节省时间。通过RT-PCR验证,2种方法所获得的草莓果实总RNA质量较好,可达到分子生物学试验对RNA质量的要求。It is difficult to extract total RNA from strawberry fruits since the fruits are rich in phenols, polysaccharide and other secondary metabolism substances. In order to establish a method for extracting total RNA from strawberry fruits, modified CTAB method and modified EASYspin plant RNA extraction kits method were compared. The concentration, integrity and purity of total RNA were detected by the agarose gel electrophoresis and nucleic acid/protein assay （NanoDrop 2000）. The results showed that both methods were fit for extracting high-quality RNA from strawberry fruits with two distinct electrophoresis bands of 28S and 18S. Both the OD260/OD230 values and the 0926o/0928o values were 2.0 above. Modified CTAB method was cheap and of high-quality, but modified EASYspin plant RNA extraction kits method was simple, safe and rapid. RT- PCR analysis showed that extracted total RNA could be applied to gene cloning and other molecular biology experiments.

关 键 词：草莓 总RNA提取 RNA提取试剂盒法 改良的CTAB法 

分 类 号：S668.4[农业科学—果树学]