SYBR Green实时定量PCR检测外源基因拷贝数  

Establishment of SYBR Green-Base Quantitative Real-Time PCR Assay for Determining Transgene Copy Number in Genome

作　　者：庄强[1] 钱程[1] 刘立[1] 

机构地区：[1]浙江理工大学生命科学院,杭州310018

出　　处：《浙江理工大学学报（自然科学版）》2010年第1期125-129,共5页Journal of Zhejiang Sci-Tech University(Natural Sciences)

基　　金："863"项目(2006AA02Z126);973计划(2004CB518804);浙江省科技厅重点项目(2006C23006)

摘　　要：以SYBR Green为荧光染料,通过携带双基因GFP和IL-24的质粒标准品绘制双标准曲线,并以绝对定量的方法检测GFP在稳定细胞株Hela-GFP基因组中的整合拷贝数。结果证实,该方法能有效检测外源基因在基因组上的整合拷贝数。从而建立起一种高效且廉价的检测外源基因拷贝数的通用方法来广泛应用于各种实验。An economical SYBR Green-base qPCR assay is established to determine copy number of GFP integrated in stable Hela-GFP cell line. Firstly, a plasmid containing GFP and IL-24 is constructed as a standard. The standard curves based on GFP and IL-24 primer are generated respectively to quantitative the total copy number of GFP and IL-24 in Hela-GFP genome sample. Then the copy number of integrated GFP can be counted with endogenous copy number of IL-24 and the ratio between them. From the result of this study, it can be concluded that this assay works well in determination of transgene copy number.

关 键 词：实时荧光定量PCR SYBR Green 整合拷贝数 

分 类 号：Q789[生物学—分子生物学]