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Digital PCR

Digital PCR Using the OpenArray® Real-Time PCR System

Digital PCR is a new approach to nucleic acid detection and quantification, which is a different method of absolute quantification and rare allele detection relative to conventional qPCR.

Digital PCR works by partitioning a sample into many individual real-time PCR reactions; some portion of these reactions contain the target molecule (positive) while others do not (negative). Following PCR analysis, the fraction negative answers is used to generate an absolute answer for the exact number of target molecules in the sample, without reference to standards or endogenous controls.

The OpenArray® Real-Time PCR System enables digital PCR experiments at a scale previously unattainable—in a single day, one user can generate >36,000 digital PCR data points on the OpenArray® Real-Time PCR System, without the use of robotics. Other features of the system include:

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OpenArray® Digital PCR Software Heatmap

Figure 1. Digital PCR uses the ratio of positive (White) to negative (Black) PCR reactions to count the number of target molecules .

  • Accurate and sensitive—detect and count individual molecules to quantify viral load, gDNA, cDNA, plasmids, or next-generation sequencing libraries
  • Fast—produce up to 144 digital answers per 3 hour run
  • Flexible—enables use of your existing assays and has the capacity to test from one to 48 assay/sample dilutions per plate
  • Wide dynamic range—with as few as 64 data points per replicate group, a dilution series can easily be loaded into the TaqMan® OpenArray® Digital PCR Plate, expanding the range of sample concentrations which can be analyzed to produce a digital answer.
  • Intuitive and economical—software includes a Poisson calculator to design your individual digital PCR experiment, eliminating optimization time and minimizing sample usage

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  • dPCR for Absolute Quantitation


    dPCR for Absolute Quantitation


    Digital PCR is quantitative PCR method that can be used to measure absolute quantitation. In this technique, the number of positive and negative amplification reactions is used to the determine precise measurement of target concentration.

    Applications
    • Absolute Quantification of Viral Load
    • Absolute Quantification of Nucleic Acid Standards
    • Absolute Quantification of Next-Gen Sequencing Libraries
    • Rare Allele Detection
    • Low-Fold Copy Number Discrimination
    • Enrichment and Separation of Mixtures
    Summary Advantages of Digital PCR:
    • No need to rely on references or standards 
    • Desired precision can be achieved by increasing total number of PCR replicates
    • Highly tolerant to inhibitors
    • Capable of analyzing complex mixtures
    • Unlike traditional qPCR, digital PCR provides a linear response to the number of copies present to allow for small fold change differences to be detected
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