Challenges for the Characterization of Genetically Modified Animals by the qPCR Technique in the Era of Genomic Editing  

作　　者：Ribrio Ivan Tavares Pereira Batista Darcio Italo Alves Teixeira Vicente Jose de Figueiredo Freitas Luciana Magalhaes Melo Joanna Maria Goncalves Souza-Fabjan 

机构地区：[1]Laboratorio de Fisiologia e Controle da Reproducao,Faculdade de Veterinaria,Universidade Estadual do Ceara(UECE),Av.Dr.Silas Munguba 1700,Fortaleza,CE,60740-903,Brazil [2]Faculdade de Veterinaria,Universidade Federal Fluminense(UFF),Rua Vital Brazil Filho,64,Niteroi,RJ,24230-340,Brazil [3]Molecular Genetics Research Unit,University Center Fametro(Unifametro),Rua Conselheiro Estelita,500,Fortaleza,CE,60010-260,Brazil

出　　处：《Veterinary Science Research》2021年第1期1-11,共11页兽医科学研究（英文）

摘　　要：Characterization of genetically modified organisms through determina­tion of zygosity and transgene integration concerning both copy number and genome site is important for breeding a transgenic line and the use of these organisms in the purpose for which it was obtained.Southern blot,fluorescence in situ hybridization or mating are demanding and time-consuming techniques traditionally used in the characterization of transgenic organisms and,with the exception of mating,give ambiguous results.With the emergence of the real-time quantitative PCR technology,different applications have been described for the analysis of transgenic organisms by determination of several parameters to transgenic analysis.However,the accuracy in quantitation by this method can be influenced in all steps of analysis.This review focuses on the aspects that influence pre-analytical steps(DNA extraction and DNA quantification methods),quantification strategies and data analysis in quantification of copy num­ber and zygosity in transgenic animals.

关 键 词：Absolute quantification Copy number DNA extraction Relative quantification ZYGOSITY 

分 类 号：Q78[生物学—分子生物学]