样本保存液和超低温冰箱保存对RT-PCR实验结果的影响  

Effects of RNAstore reagent and ultra-low temperature freezer on the results of RT-PCR test

作　　者：王静[1] 庞晓楠[1] 林爱琴[1] 李铁臣[1] 

机构地区：[1]皖南医学院分子生物学研究室

出　　处：《皖南医学院学报》2013年第6期436-438,共3页Journal of Wannan Medical College

基　　金：安徽高校省级自然科学研究重点项目(KJ2010A321)

摘　　要：目的:通过对两种新鲜组织保存方法的比较,探讨一种简便、有效的用于抽提RNA的保存新鲜组织的方法。方法:采集人新鲜子宫内膜组织标本38例,其中26例组织用样本保存液(RNAstore Reagent)常温保存,12例组织冻存于-80℃冰箱,1周后提取总RNA。β-Actin基因mRNA的表达水平通过两种方法检测。一种经1.0%琼脂糖凝胶电泳,溴化乙锭染色,紫外透射仪下观察分析,另一种是使用ABI PRISM 7500 PCR仪进行荧光定量PCR检测。比较两组间β-Actin基因的光密度值和平均CT值的差异。结果:两种方法保存的新鲜组织标本均出现305 bp的目标带。OD值分别为2 368.24±678.01和2 493.35±696.181,平均CT值分别为13.64±3.31和14.08±1.57,两组之间无显著性差异(P>0.05)。结论:在没有超低温冰箱的情况下,样本保存液(RNAstore Reagent)可以用来保存新鲜组织样本。Objective：To probe into a simple and effective approach to preserving fresh tissue specimen for extracting RNA by comparing two di-verse techniques.Methods： Fresh endometrial tissues were procured from 38 individuals and stored either in RNAstore reagent（26）or in icebox at-80℃（ 12 ） , respectively.Total RNA was directly extracted after one week with Trizol reagent, and expression levels of β-Actin gene mRNA were detected with two different methods .One group of specimens were treated with 1.0% agarosegels electrophoresis and ethidium bromide stai-ning before ultraviolet transilluminator detection , and another group were undergone direct fluorescent quantitation using ABI PRISM 7500 real-time PCR device for comparing the optical density value （ OD value） and average CT value of β-Actin gene between the two groups.Results：Generally,two preservation techniques resulted in visual target band at 305 bp.The OD values of β-Actin were 2 368.24 ±678.01 and 2 493.35 ±696.18,and the average CT values of β-Actin were 13.52 ± 3.4 and 14.08 ±1.57,respectively.No significant difference was found between the two methods used（P 〉0.05）.Conclusion： The findings suggest that RNAstore reagent can be used for preserving fresh tissue specimens in case ultra low temperature freezer were not available .

关 键 词：子宫内膜组织 样本保存液 RNA提取 RT-PCR QPCR 

分 类 号：R446.1[医药卫生—诊断学]