避免基因组DNA污染的半定量RT-PCR新方法  

A New Semiquantitative RT-PCR Technique to Avoid Genome DNA Contamination

作　　者：卢加举[1] 崔百明[2] 廖文彬[2] 于海川[2] 彭明[2] 

机构地区：[1]贵州省亚热带作物研究所,贵州兴义562400 [2]中国热带农业科学院热带生物技术研究所,海南海口571101

出　　处：《天津农业科学》2011年第6期6-9,共4页Tianjin Agricultural Sciences

基　　金：贵州省科学技术基金(黔科合J字[2008]2077);国家973项目(2004CB117307)

摘　　要：为了消除基因组DNA的污染,在逆转录引物oligo-dT5’端加上通用引物T7B结合序列,根据3’-RACE的原理,用T7B和目标基因特异性引物即可进行半定量RT-PCR分析。用该方法对转GhGAI基因的拟南芥进行表达分析,结果表明:此方法可有效避免基因组DNA污染导致的假阳性。In this paper,a new semi-quantitative reverse transcription PCR technique was employed for analyzing the expression of genes,which have no intron or unknown intron.To deal with the existence of genome DNA contamination,universal primer T7B link sequence was added to oligo-dT-5＇end.The reverse transcription PCR was performed by using T7B and special primer of target according to 3 ＇RACE theory.The expression of GhGAI gene in transgenic Arabidopsis was analyzed using this method,the results indicated that this method could effectively remove genome DNA contamination and prevent against false-positive result.

关 键 词：半定量RT-PCR 新方法 转基因植株 

分 类 号：X171[环境科学与工程—环境科学]