一种基于双重PCR结合毛细管电泳验证转录组测序结果的新方法  

Combination of duplex PCR technology with capillary electrophoresis is an alternative method to validate the result of transcriptome sequencing

作　　者：易健明 高艳[2] 李志慧[2] 张艳春[2] 屈武斌[2] 张成岗[1,2] 

机构地区：[1]安徽医科大学研究生院,合肥230032 [2]北京放射与辐射医学研究所、蛋白质组学国家重点实验室、全军军事认知与心理卫生研究中心,北京100850

出　　处：《军事医学》2014年第9期724-728,共5页Military Medical Sciences

基　　金：国家重点基础研究发展计划资助项目(973计划)(2012CB518200);国家自然科学基金资助项目(31101049,31371345,31300703);国家863计划项目(2012AA022402);航天医学基础与应用国家重点实验室2011年开放课题(SMFA12K15);中国科学院海洋生物资源可持续利用重点实验室2011年开放课题(LMB111001)

摘　　要：目的将双重PCR与毛细管电泳技术相结合,建立一种验证转录组测序结果的新方法。方法根据前期进行转录组测序的分析结果,挑选8个差异基因作为靶基因,分别使用双重PCR结合琼脂糖凝胶电泳、双重PCR结合毛细管电泳以及实时荧光定量PCR进行验证,比较三者的验证率。结果双重PCR结合琼脂糖凝胶电泳的验证率为50%;双重PCR结合毛细管电泳和实时荧光定量PCR的验证率均为100%。结论双重PCR结合毛细管电泳可作为转录组测序结果验证的一种有效方法。Objective To establish a method to validate the result of transcriptome sequencing using duplex PCR technology combined with capillary electrophoresis. Methods According to a previous study on transcriptome sequencing,eight differentially expressed genes were chosen as target genes for examination. The mRNA expression level of these genes was detected using duplex PCR combined with agarose gel electrophoresis,duplex PCR combined with capillary electrophoresis and Q-PCR,respectively. Then,the verification efficiency of each method was evaluated carefully. Results The verification efficiency of duplex PCR combined with agarose gel electrophoresis was 50%,while that of duplex PCR combined with capillary electrophoresis and Q-PCR was both 100%. Conclusion Combination of duplex PCR technology with capillary electrophoresis can be used as an alternative method to validate the results of transcriptome sequencing.

关 键 词：转录组测序 双重PCR 毛细管电泳 实时荧光定量PCR 

分 类 号：Q503[生物学—生物化学]