芍药芽总RNA提取方法的研究  

Study on Extraction Method of Total RNA from Peony Buds

作　　者：胡小蓉[1] 马燕[1] 臧德奎[1] 宋淑香[1] 郭先锋[1] 

机构地区：[1]山东农业大学林学院,山东泰安271018

出　　处：《山东农业科学》2015年第2期16-20,共5页Shandong Agricultural Sciences

基　　金：山东省农业良种工程重大课题[鲁农良字(2010)6号]

摘　　要：以处于休眠和解除休眠时期的芍药芽为试验材料,采用改良CTAB法、Trizol法和EASYspin Plus植物RNA快速提取试剂盒提取其总RNA,通过核酸蛋白仪、琼脂糖凝胶电泳和RT-PCR检测所提总RNA样品的品质。结果表明,Trizol法没有提取到芍药芽总RNA;EASYspin Plus植物RNA快速提取试剂盒提取的总RNA浓度太低,无法满足下一步试验的要求;改良CTAB法提得的总RNA较完整,条带清晰,品质较优,28S rRNA亮度约是18S rRNA的2倍,且无降解现象。以改良的CTAB法提取的总RNA为模板进行内参Actin扩增,得到的条带清晰,与目的片段大小一致,进一步证明此方法提取的总RNA能够满足后续试验的要求。The total RNA was extracted from dormancy and dormancy -released peony buds by modified CTAB method, Trizol method and EASYspin Plus RNA rapid isolation kit respectively. The quality of the ex- tracted RNA was tested by protein nucleic acid analyzer, agarose gel electrophoresis and RT -PCR. The re- sults showed that the total RNA could not be extracted by Trizol method, and that extracted by EASYspin Plus RNA rapid isolation kit was too lower to meet the requirement of further experiment. The total RNA extracted by modified CTAB method was more complete with clearer bands and better quality, and the brightness of 28S rRNA was about 2 times of that of 18 S rRNA with no degradation. Using the total RNA extracted by modified CTAB method as template to amplify the internal reference gene Actin, the obtained bands were clear and with the same size to the target fragment, which further proved that the total RNA extracted by modified CTAB method could meet the requirement of the later experiment.

关 键 词：芍药芽 总RNA 提取 RT-PCR 

分 类 号：S682.120.1[农业科学—观赏园艺]