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Bidirectional Retroviral Integration Site PCR Methodology and Quantitative Data Analysis Workflow

Article DOI: 10.3791/55812-v • 12:53 min • June 14th, 2017
June 14th, 2017 •
*1,2, *3, 1, 3, 4, 1,5, 6
1UCLA AIDS Institute, University of California at Los Angeles (UCLA), 2Department of Microbiology, Immunology, & Molecular Genetics, University of California at Los Angeles (UCLA), 3Departments of Biomathematics and Mathematics, University of California at Los Angeles (UCLA), 4Personalized Genomic Medicine Research Center, Division of Strategic Research Groups, Korea Research Institute of Bioscience and Biotechnology, 5Department of Medicine, University of California at Los Angeles (UCLA), 6Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University (OSU)
* These authors contributed equally

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This manuscript describes the experimental procedure and software analysis for a bidirectional integration site assay that can simultaneously analyze upstream and downstream vector-host junction DNA. Bidirectional PCR products can be used for any downstream sequencing platform. The resulting data are useful for a high-throughput, quantitative comparison of integrated DNA targets.

Tags

Retroviral Integration Site PCR Quantitative Data Analysis Retroviral Vector Integration Sites Clonal Cell Populations Host Genome Gene Therapy Genetically Engineered Cell Retro-integration Site Sequences DNA Concentration Absorbance Ratio Genomic DNA Analysis PCR Reaction Biotin Primers Lentivirus-specific
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