2种方法从大鼠晶状体中提取总RNA及评测  

Evaluation of Two Methods for Total RNA Extraction in Rat Lens

作　　者：徐洪卫 张晨光[1] 金磊[2] 

机构地区：[1]徐州医学院临床学院,江苏徐州221004 [2]徐州医学院医学生物化学与分子生物学教学实验中心,江苏徐州221004

出　　处：《安徽农业科学》2014年第35期12423-12424,12428,共3页Journal of Anhui Agricultural Sciences

基　　金：江苏省大学生创新计划资助项目(2012JSSPITP1823)

摘　　要：[目的]测评2种方法从大鼠晶体中提取总RNA的效果[方法]采用2种方法从大鼠晶状体中提取总RNA,通过测定总RNA的浓度以及A260/A280、A260/A230、琼脂糖凝胶电泳、RT-PCR方法对其进行评价.[结果]使用柱式法从大鼠晶状体中提取总RNA,其28S、18S亚基清晰可见,NanoDrop22000测定显示A260/A280、A260/A230均在1.8 ～2.0,可用于反转录聚合酶链反应等下游试验.Trizo1法提取结果显示总RNA降解较多,不适合进行下游试验.[结论]柱式法操作简便,全程可在常温下进行,总RNA质量高,适宜于实验教学及科研.Trizol法提取总RNA操作要求相对较高,且易受到苯酚污染及乙醇残留,但适合抽提大量样本的总RNA,对于实验教学有一定难度.[Objective] The aim was to make evaluation of two methods for total RNA extraction in rat lens.[Method] Two methods were used to extract total RNA from the rat lens.And it were evaluated by measuring A260/A280,A2600/A230,agarose gel electrophoresis,and RT-PCR method.[Result] Results showed that using column method to extract total RNA from the rat lens,the 28S and 18S subunits were clearly visible.Both A260/A280 and A260/A280 values were between 1.8 and 2.0.The results could be used in the downstream experiments such as reverse transcription polymerase chain reaction.Using Trizol method,the results showed that the total RNA was degenerated.The experiment results were not suitable for the downstream experiment.[Conclusion] Column method that was simple,and could operate at room temperature and obtain the high quality of total RNA was suitable for experiment teaching and scientific research.Trizol method of extracting total RNA operating requirements were relatively high,and susceptible to phenol pollution and residual ethanol,but suitable for total RNA extraction of sample and had the certain difficulty for experimental teaching.

关 键 词：总RNA 柱式法 TRIZOL法 

分 类 号：S188[农业科学—农业基础科学] Q-3[生物学]