不同沉淀方法处理胶乳总RNA对基因扩增效率的影响  

Effects of the Latex Total RNA Precipitated with Different Methods on Amplification Efficiency

作　　者：吴春太[1] 黎瑜[1] 冯伟瑜 曾日中[1] 

机构地区：[1]中国热带农业科学院橡胶研究所农业部橡胶树生物学与遗传资源利用重点实验室海南省热带作物栽培生理学重点实验室,儋州571737 [2]海南大学农学院,海口570228

出　　处：《生物技术通报》2016年第11期124-129,共6页Biotechnology Bulletin

基　　金：国家自然科学基金资助项目(31270713);橡胶研究所基本科研业务费资助项目(1630022012008)

摘　　要：为探明沉淀方法对胶乳总RNA提取及PCR扩增效率的影响,并寻求较好的RNA沉淀方法,针对胶乳中蛋白、多糖、橡胶粒子含量高的特点,在本实验室改进的SDS法的基础上,分别用Li Cl、Na Cl+异丙醇、Na Cl+Na Ac+无水乙醇和Na Ac+无水乙醇对橡胶树胶乳总RNA进行沉淀,并对所得RNA的完整性、纯度、含量和RT-PCR扩增效率进行比较。结果表明,4种方法均可获得完好的RNA,但其纯度、含量和PCR扩增效率随方法不同而有所差别,其中Na Cl+Na Ac+无水乙醇沉淀的橡胶树胶乳总RNA纯度、产率和18S、REF基因扩增效果均较高,故认为用Na Cl+Na Ac+无水乙醇沉淀胶乳总RNA是一种较为有效的方法。This work is to find out the effects of different precipitation methods on the latex total RNA and amplification efficiencyof PCR as well as to seek the better precipitation method. According to the characteristics that Herva brasiliensis latex was rich in proteins,polysaccharides and rubber particles,on the basis of SDS method modified in our laboratory,the total RNA in the latex of H. brasiliensiswas precipitated respectively with Li Cl,Na Cl ＋ isopropanol,Na Cl ＋ Na Ac ＋ absolute ethanol,and Na Ac ＋ absolute ethanol,and then theintegrity of the RNA,purity,content,and RT-PCR amplification efficiency were compared. The results indicated that complete RNA wasobtained by any of 4 methods. But the differences in purity and content of RNA and amplification efficiency of RT-PCR among 4 methods werealso observed. In which the purity and yield of total RNA precipitated with Na Cl ＋ Na Ac ＋ absolute ethanol and 18 S and REF gene amplificationefficiency of latex tissues in rubber tree were higher,indicating that Na Cl ＋ Na Ac ＋ absolute ethanol was effective method to precipitate totalRNA.

关 键 词：橡胶树胶乳 总RNA提取 RNA沉淀 

分 类 号：S794.1[农业科学—林木遗传育种]