桃不同发育时期叶片总RNA提取方法的比较  

Comparative Analysis of Total RNA Extraction Methods for Leaves of Peach at Different Development Stages

作　　者：王其海[1] 王裔娜[1] 刘晓华[1] 吴文江[2] 丁宁[1] 牛洪斌[3] 王林忠[1] 吴国良[1] 

机构地区：[1]河南农业大学园艺学院,河南郑州450002 [2]河南农业大学食品科技学院,河南郑州450002 [3]河南农业大学国家小麦工程技术研究中心,河南郑州450002

出　　处：《河南农业科学》2014年第3期116-120,共5页Journal of Henan Agricultural Sciences

基　　金：国家自然科学基金项目(31171943);河南省重大科技攻关项目(92101110600)

摘　　要：为了从桃叶片中提取到高质量的RNA,以进行反转录、RT-PCR等后续分子生物学试验,选取了改良硼砂-CTAB法、改良CTAB法、改良SDS-苯酚法、试剂盒法、Trizol法,以及在试验中探索的改良Trizol法Ⅰ和改良Trizol法Ⅱ对桃不同发育时期的叶片总RNA进行提取,测定RNA样品的OD值,结合琼脂糖凝胶电泳检测总RNA样品的质量。结果表明,这些方法虽然都能获得桃叶片总RNA,但是或存在不同程度的DNA残留,或有蛋白、多糖等杂质污染,或降解严重;而改良的Trizol法提取总RNA的完整性和纯度较好;针对生长期和衰老期叶片的不同生理特性可分别采用改良Trizol法Ⅰ和改良Trizol法Ⅱ,提取的总RNA反转录后,通过RT-PCR检测,条带明亮清晰,与预期目的基因片段大小一致,说明这2种方法提取的总RNA能用于后期的分子生物学试验。The total RNA of peach leaves at different development stages was extracted by the improved borax-CTAB method,improved CTAB method, improved SDS-phenol method, kit method, Trizol method, and the improved Trizol I and Trizol II method,then the purity and concentration of the total RNA sample was determined. The results showed that the improved Trizol methods were better;for the other methods,there were different degrees of DNA residues, or pollution, or degradation;improved Trizol I and Trizol II method were better for the extraction of total RNA from peach leaves at vegetative stage and aging stage, respectively, and the total RNA extracted by the improved Trizol methods met the needs of the follow-up molecular biological study.

关 键 词：桃叶片 RNA提取 TRIZOL法 改良CTAB法 改良SDS-苯酚法 

分 类 号：S662.1[农业科学—果树学]