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Comparing ddPCR with qPCR for the quantification of AAV vector genomes

Bio-Rad
7 Apr 2022

Developing effective and reproducible gene therapies requires the use of sensitive and robust testing methods, such as Droplet Digital PCR, to validate the quality of the therapeutic product. Bio-Rad's portfolio of ddPCR Viral Quantification solutions offers an accurate, precise, and robust method to measure the viral titer in a sample.

In this whitepaper, Martin Lock, Director, Process Development Gene Therapy Program, University of Pennsylvania, shares data comparing an AAV genome titer assay based on ddPCR with a standard and an optimized qPCR assay for the titration of both single-stranded (ss) and self-complementary (sc) AAV genomes.

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ddPCR Cell and Gene Therapy Assays

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Bio-Rad’s ddPCR Cell and Gene Therapy (CGT) Assays are custom ddPCR-optimized assays generated by the Cell and Gene Therapy Design Engine for your target Gene of Interest. These assays offer the ease and accuracy of ddPCR technology and come in multiple fluorophores to enable multiplexed Cell and Gene Therapy applications, including Viral Titer and Vector Copy Number measurements. Use the ddPCR CGT Assays individually or multiplex them with the Vector Backbone Assays to precisely measure the concentration of the therapeutic gene in your sample. Features and Benefits Quick assay design through the Cell and Gene Therapy Design Engine, saving you valuable time to focus on important experiments Robust assay design, optimized specifically for ddPCR technology Six probe fluorophore options to enable multiplexing Vector Backbone Assays available for multiplexing In-silico multiplexing checks (wet-lab validation recommended) Certificate of Analysis provided with each ddPCR CGT Assay order Assay sequence provided with select catalog numbers Applications and Uses Viral Vector Genome Titer Measurement Vector Copy Number Determination Gene Therapy Development and Manufacturing CAR-T Cell Therapy Development and Manufacturing

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QX200™ Droplet Digital PCR System

Bio-Rad

Droplet Digital PCR System, includes droplet generator, droplet reader, laptop computer, software, associated component consumables, for EvaGreen or probe-based digital PCR applications

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PCR and Thermal CyclingPolymerase chain reaction (PCR) kits and thermal cyclers are used for the in vitro amplification of DNA permitting subsequent analysis and experimental procedures. Explore a range of high-quality polymerase, primers and nucleotides or simplify your workflow with a PCR mastermix. Find reverse transcription PCR (RT-PCR) and cDNA synthesis kits for RNA products and libraries. Quantitatively measure the amplification of DNA with real-time PCR (qPCR) and droplet digital PCR (ddPCR) kits and systems, and discover automated PCR setup solutions to increase throughput. Alternative DNA amplification methods also include recombinase polymerase amplification (RPA) kits. Find the best PCR kits and thermal cyclers and purification equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Drug DeliveryDrug Delivery refers to dosage form, route of administration, formulations, technologies, and systems for transporting a pharmaceutical compound in the body as needed to safely achieve its desired therapeutic effect. Drug delivery is often approached via the biopharmaceutical or small molecule drug's formulation, but it may also involve medical devices or drug-device combination products. Considerations include instrumentation, software and services. Bioprocessing / FermentationBioprocessing is the use of biological materials to perform commercial, scientific or medical research processes. Biological materials used include cells, enzymes and organisms. Usually bioprocessing requires a batch or continuous bioreactor such as a fermentor or cell culture system. The advantages of using a reactor include high productivity, easy configuration, adjustable values and automation.DNA / RNA QuantificationDetection and quantification of nucleic acids is important in molecular biology, cloning, expression, forensics and clinical diagnostics. Nucleic acids can be detected by labeling with colorimetric, fluorescent or radio labels and using in situ hybridization kits to identify specific sequences. Multiple nucleic acids can be detected and quantified at once using RNA / DNA detection beads or RNA / DNA microarrays. Find the best DNA / RNA Quantification products in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Droplet Digital PCRDroplet digital PCR (ddPCR) is a method of quantitative PCR that partitions a sample into droplets, allowing precise detection of low-abundance targets. It is commonly used in gene expression analysis and mutation detection. Explore ddPCR systems in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.qPCRQuantitative PCR (qPCR) is a molecular biology technique used to amplify and measure the amount of a specific DNA or RNA target. It plays a crucial role in gene expression analysis, pathogen detection, and quantifying DNA or RNA in various applications. Real-time PCR provides precise, quantitative results and is widely used in research, diagnostics, and clinical studies. Browse our peer-reviewed product directory to find the best qPCR solutions; compare products, check reviews, and get pricing directly from manufacturers.DNA QuantificationDNA quantification is the process of measuring the amount of DNA in a sample. It is vital in genomics, molecular biology, and biotechnology, helping to ensure accurate experiments and data analysis. Explore DNA quantification tools in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.VirusesViruses are microscopic pathogens that require a host cell to replicate. Understanding their structure, replication cycle, and impact on the immune system is crucial in developing effective treatments, vaccines, and diagnostic methods. Research continues to focus on emerging viral diseases and antiviral drug discovery. Explore solutions for virus research, detection, and treatment in our peer-reviewed product directory; compare products, check customer reviews, and get pricing directly from manufacturers.

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