Should You Use Digital or Real-Time PCR?

digital-real-time-pcr-99808
digital-pcr

Overview

Absolute measurement: Uses single-molecule amplification to count the target of interest across a large number of PCR replicates; use limiting dilution to ensure at least one reaction does not contain the target.

real-time-pcr

Overview

Uses a reference to measure PCR amplification as it occurs; data collected during the exponential (log) phase of PCR.

Applications

  • Rare target detection
  • Absolute quantitation of gene expression
  • Absolute quantitation of viral load
  • Absolute quantitation of nucleic acid
  • References and standards

Applications

  • SNP genotyping
  • Relative gene expression analysis
  • MicroRNA analysis
  • Standard copy number variation

Advantages

  • No need for references or standards
  • Increased performance by increasing total number of PCR replicates
  • Highly inhibitor-tolerant
  • Can analyze rare targets against wild-type background
  • Provides linear response to the number of copies; allows for small fold-change differences to be detected

Advantages

  • Broadly accepted, well-established protocols and assays
  • Dynamic range of detection
  • Detection down to a two-fold change
  • High sample throughput, lower cost
  • Data collected during the exponential phase of PCR; provides a permanent record of amplicon amplification with the cleaved probe