Digital PCR is a new approach to nucleic acid detection and quantification. It offers a different method for absolute quantification and rare allele detection relative to conventional real-time quantitative PCR. 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 of negative reactions is used to generate an absolute count of the number of target molecules in the sample, without reference to standards or endogenous controls.

Applications Advantages of Digital PCR
  • Rare allele detection
  • Absolute quantification of gene expression
  • Absolute quantification of viral load
  • Absolute quantification of nucleic acid standards
  • Absolute quantification of next-generation sequencing libraries
  • No need to rely on references or standards
  • Desired precision can be achieved by increasing the number of PCR replicates
  • Highly tolerant to inhibitors
  • Capable of analyzing complex mixtures
  • Provides a linear response to the number of copies present to to help enable analysis of small fold-change differences

The GRCF offers digital PCR using the LifeTechnologies QuantStudio 3D system.  This is not a droplet based system.  Instead, the sample is spread onto chips, partitioning the sample into about 18,000 individual wells.  The system uses Taqman assays or assays labelled with Fam or Vic.



*ThermoFisher recommends running 2 chips per sample.