影响RNA提取质量的因素分析  

Analysis of factors affecting RNA isolation qualities

作　　者：宗成志[1] 左欣欣[1] 严海燕[1] 

机构地区：[1]中南民族大学生命科学院,湖北武汉市430073

出　　处：《广西农学报》2011年第1期31-34,共4页Journal of Guangxi Agriculture

基　　金：中南民族大学校基金资助项目:YZZ06004

摘　　要：RNA提取技术已经成为分子生物学中一个基本技术。教学和有一定特殊要求的科学研究需要透彻了解影响RNA提取的详细因素。细节对RNA提取的质量至关重要。本文对CTAB法提取RNA的全过程中各种影响因子进行了分析:CTAB法可以被用于RNA的提取;在CTAB提取缓冲中的温育时间以及机械摇动力是影响所提RNA质量的两个主要因素;合适的温育时间是65℃5min;轻柔地将组织与CTAB提取缓冲液充分混合可以避免DNA的污染;采用此方法进行实验可在在酚抽提的步骤停下,同时不影响RNA的质量,给实验安排带了灵活性。这是一个用于一般实验室的简单便宜的方法。RNA isolation has been a basic technology of molecular biology.In the lab teaching,detailed factors affecting RNA extraction need to be understood fully,in which detailed condition is critical for quality of extracted RNA.In this paper,various factors affecting RNA extraction quality with CTAB method were studied.The results show that Poly VinylPyrrolidone and spermidin can increase RNA output,incubation time in CTAB for 5 mins is enough and suitable for good quality of RNA,strong mechanical shaking can cause DNA contamination,chloroform-isoamyl alcohol（24∶1） extraction after phenol extraction is necessary for clearing of phenol comtamination in RNA.With a 5 mins CTAB extraction,subsequently with phenol,phenol/chloroform-isoamyl alcohol,chloroform-isoamyl alcohol extraction and precipitation with isopropanol procedure,RNA can be extracted with pure and good quality for reverse transcription and QPCR.This is also a simple and unexpensive method for general lab.

关 键 词：RNA提取 聚乙烯基吡咯烷酮 十六烷基三甲基溴化铵 加热时间 酚/氯仿—异戊醇 

分 类 号：Q522.03[生物学—生物化学]