灰毡毛忍冬花蕾总RNA提取方法的研究  

Study on Methods for Extraction of Total RNA from Flower Bud of Lonicera Macranthoides

作　　者：徐玉琴[1] 王珊[1] 刘湘丹[1] 童巧珍[1] 周日宝[1] 孙梦姗[1] 

机构地区：[1]湖南中医药大学,湖南长沙410208

出　　处：《中医药导报》2015年第24期31-34,共4页Guiding Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金(81203007);国家中医药管理局"药用植物学"重点学科资助(No.国中医药发[2009]30号);湖南省自然科学基金(13JJ9010);湖南省教育厅科技创新平台(14K071);湖南省"中药学"重点学科建设项目资助(No.湘教通[2011]76号)

摘　　要：目的:探讨灰毡毛忍冬花蕾总RNA提取的有效方法。方法:采用SDS法、Trizol法、普通试剂盒法、改良CTAB法、多糖多酚试剂盒法、改良多糖多酚试剂盒法提取灰毡毛忍冬花蕾总RNA并比较其效果。结果:SDS法、Trizol法和普通试剂盒法均不适用于本样品总RNA的提取;改良CTAB法、多糖多酚试剂盒法、改良多糖多酚试剂盒法均能有效去除蛋白质、多糖多酚及DNA,OD260/280为1.86-2.00,OD260/230为1.96-2.63,电泳条带清晰,RNA完整性好,且通过RT-PCR扩增能获得特异性条带。但以多糖多酚试剂盒法提取效果最佳。结论:改良CTAB法、多糖多酚试剂盒法、改良多糖多酚试剂盒法均适合灰毡毛忍冬花蕾总RNA提取,完全满足后续RT-PCR等分子生物学研究。Objective： To explore a method for extraction of the total RNA from flower bud of Lonicera macranthoides. Methods： By comparing 6 kinds of extraction methods, such as, SDS, Trizol, Ordinary kit method, improved CTAB, Polysaccha- ride polyphenols kit method and improved Polysaccharide polyphenols kit method to isolate total RNA from flower bud of Lonicera macranthoides. Result： SDS, Trizol and Ordinary kit method were not suitable for this experiment; Improved CTAB, Polysaccharide polyphenols kit method and improved Polysaccharide polyphenols kit method could isolate total RNA, which OD260/280 was 1.86-2.00, OD260/230 was 1.96-2.63, and had less protein, DNA and polyphenol polysaccharide pollution. The electrophoresis bands were cleared on agarose gel and integrity of RNA was good. The RT-PCR amplified specific bands also can be obtained. Polysaccharide polyphenols kit method is the best method. Conclusion： Improved CTAB, Polysaccharide polyphenols kit method and improved Polysaccharide polyphenols kit method all can be used to isolate the total RNA from flow- er bud of Lonicera macranthoides, which can completely meet the following related experiment.

关 键 词：灰毡毛忍冬 花蕾 总RNA提取 改良CTAB法、多糖多酚试剂盒法 

分 类 号：R284.2[医药卫生—中药学]