银杏不同组织的总RNA提取方法的改进  

Modification of Total RNA Isolation Method from Different Ginkgo biloba Organs

作　　者：蔡荣[1] 许锋[1] 陈柳吉[1] 程水源[1] 

机构地区：[1]长江大学园艺园林学院,湖北荆州434025

出　　处：《生物技术》2007年第4期38-41,共4页Biotechnology

基　　金：教育部新世纪优秀人才计划项目资助(NCET-04-0746);教育部地方重点项目(02095);湖北省自然科学基金项目资助(2002AB094);湖北省青年杰出人才基金项目(2003AB014);湖北省教育厅重大科技项目(Z200627002)资助

摘　　要：方法:采用改进的CTAB法高效地从富含多糖、多酚类化合物的银杏的根、茎、叶和果实四个组织中提取RNA。结果:抽提的不同组织的RNA经电泳检测,可见28S和18S两条清晰的主带,且28S rRNA在亮度上均为18S rRNA的2倍,两条带之间无弥散现象;根、茎、叶、果所提的RNA的经紫外吸收检测得到A260/A230分别为1.9、1.4、2.1、1.7,测得A260/A280分别为1.8、2.0、2.3、1.8,鲜重分别达到78μg/g、69μg/g、150μg/g、90μg/g;通过RT-PCR及凝胶检测,得到了银杏看家基因18S基因的约150bp清晰的条带,从而进一步检测提取RNA的质量。结论:提取的总RNA纯度高,质量好,足以为研究提供RNA材料。An improved CTAB extract protocol was developed to isolate total RNA in good yield and integrity from Ginkgo biloba mot, stalk, leaf and fruit containing high levels of carbohydrates and polyphenolic secondary metabolites weight. At least two distinct 28S and 18S rRNA distinct major bands were detected by denaturing gel electrophoresis, and the sharpness of 28S bands was twice than 18S of bands . A260/A230 ultraviolet radiation absorbancy ratios of RNA isolate from Ginkgo biloba root, stalk, leaf and fruit were 1.9,1.4,2.1,1.7 and A260/A280 ultraviolet radiation absorbancy ratios were 1.8,2.0,2.3,1.8 and the yield were 78μg/g ,69μg/g, 150μg/g ,90μg/g flesh weight. A Cinkgo biloba house keeping gene fragment： the 150 bp sequence of 18S gene was successfully amplified by RT- PCR, suggesting the integrity of isolated RNA. The total RNA isolated by this protocol is of sufficient quality for subsequent molecular applications.

关 键 词：银杏 不同组织 总RNA提取 

分 类 号：Q781[生物学—分子生物学]