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  11. Digital PCR Using the OpenArray® Real-Time PCR System

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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