长距离RT-PCR非特异性扩增的鉴定与排除  

Identification of Non-specific Amplification with Long RT-PCR

作　　者：唐青海[1] 乔宪凤 何维明[1] 郑新民 华文君 刘西梅 陈果亮[3] 马秋明[1] 

机构地区：[1]西北农林科技大学动物科技学院,陕西杨凌712100 [2]湖北省畜牧兽医研究所,湖北武汉430064 [3]扬州大学兽医学院农业部畜禽传染病重点实验室,江苏扬州225009

出　　处：《安徽农业科学》2007年第15期4448-4450,共3页Journal of Anhui Agricultural Sciences

基　　金：湖北省自然科学基金项目(2005ABA195)

摘　　要：根据已经测定的乙脑病毒WHe株全基因组序列(GenBank EF107527),设计1对引物扩增基因组全长cDNA和2对特异性的鉴定引物。经RT-PCR技术扩增后,得到约9 kb的产物,经过反复实验,鉴定为外源性污染核酸扩增产物,并加以有效的排除,初步的研究表明该产物可能是由外源性污染核酸自身反向部分互补扩增所致。Two primers used to amplify the full length of Japanese encephalitis virus （JET） WHe strain and another two pars of primers used for identification were designed according to the complete genome of JEV-WHe strain （GenBank number, EF107527） .A 9 kb cDNA fragment was produced with the long RT-PCR, and it was identified as a non-specific amplification. The research showed that this products was amplified from unknown DNA or RNA templates.

关 键 词：RT-PCR 乙脑病毒 非特异性扩增 

分 类 号：Q819[生物学—生物工程]