Transitioning your assay from quantitative PCR to droplet digital PCR

28 Mar 2022

In this application note, Bio-Rad discusses migrating assays from real-time quantitative PCR (qPCR) to Droplet Digital PCR (ddPCR). These two PCR methods determine the concentration of sample DNA using fundamentally different approaches and a bridging study between the two methods presents data demonstrating the differences and similarities to consider.

QX ONE™ Droplet Digital™ PCR System

Bio-Rad

The QX ONE Droplet Digital PCR System integrates droplet generation, thermal cycling, droplet reading, and analysis into a single automated precision platform that delivers superior precision and accuracy and absolute quantification.

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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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Cell / Tissue CultureCell culture or tissue culture is used to study the biology of cells or tissues and to isolate cellular products in an environment which can be manipulated and well defined. Accurately control your culture environment with bioreactors or culture incubators, bind your cells to a surface or together with an extracellular matrix. Distinguish cell types with differential media or proliferate cells with certain characteristics using selective media. Enrich your media with supplements such as growth factors, sera and vitamins. Find the best cell and tissue culture products, kits and equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.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.Cell Lines Stem Cells and Primary CellsPrimary cell cultures, established cell lines and stem cells are vital for <i>in vitro</i> and <i>ex vivo</i> experimentation. High-quality cells, optimized for your applications, alongside optimized cell substrates, growth medium and supplements, are critical for experimental success. Explore a range of cells suitable for your applications, including isogenic cell lines, competent cells, induced pluripotent stem cell (iPSC)-derived cell lines, fungal/bacterial/mammalian cell lines, stem cells and cancer cell lines. Find the best cells for your research in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Cell-Based AssaysCell-based assays are used to monitor the presence, quantity and activities of a desired cellular analyte including drug molecules or biomarkers. This can reveal information on cell health (apoptosis, cytotoxicity, viability and proliferation assays), cell metabolism, cell migration and cell signaling mechanisms. Find the best cell-based assay products, kits and equipment with our peer reviewed product directory: compare products, check customer reviews and receiving pricing direct from manufacturers.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.Molecular BiologyMolecular biology is the branch of biology that focuses on the molecular mechanisms that underlie cellular functions. It involves studying DNA, RNA, and proteins to understand gene expression, replication, and regulation. Molecular biology is fundamental to biotechnology, medicine, and genetic research. Explore molecular biology products 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.DNADeoxyribonucleic Acid (DNA) is the main component of chromosomes and the carrier of genetic information of living organisms. Find out here about PCR, NGS, ChIP-Seq, gel imaging, and many other techniques which can be used for the analysis of DNA.Method DevelopmentMethod development is the process of creating and optimizing experimental techniques and protocols to achieve reliable, reproducible results. This is essential in various fields, including pharmaceuticals, environmental science, and diagnostics. Browse our peer-reviewed product directory to find the best method development solutions, compare products, check reviews, and get pricing directly from manufacturers.Cell TherapyCell therapy involves using living cells to treat diseases, often by replacing damaged cells or stimulating regeneration. Stem cell therapy and CAR-T cell therapy are examples of cutting-edge treatments in regenerative medicine and cancer immunotherapy. Browse our peer-reviewed product directory to find the best cell therapy tools, compare products, check reviews, and get pricing directly from manufacturers.Gene TherapyGene therapy is a technique that modifies or replaces genes within an individual's cells to treat or prevent diseases. This approach holds potential for curing genetic disorders, cancers, and certain viral infections. Advances in gene editing technologies like CRISPR are driving the growth of gene therapy. Explore gene therapy tools and technologies in our peer-reviewed product directory; compare products, check reviews, and get pricing directly from manufacturers.
Transitioning your assay from quantitative PCR to droplet digital PCR