Conventional PCR Has Its Limitations, Even After 20 Years

Symmetric PCR has been the dominant method of DNA amplification for the last 20 years. Each amplicon in such assays is generated by use of two primers of equal concentration and equal Tm (melting temperature) and is therefore double-stranded. Amplification proceeds exponentially, but then slows down and reaches plateau in a stochastic manner. Thus, although such reactions are well known and are often robust, they nevertheless suffer from limitations which are particularly evident in samples containing low numbers of initial targets.  These limitations include:

  • Design difficulties, particularly for multiplexing
  • Reduced efficiency and sensitivity due to mis-priming
  • Variability among replicate reactions, particularly at end-point
  • Limited availability of fluorescent colors
  • Limitations in probe design
  • Requirement for hot-start enzymes
  • Costs

A Technology Platform That Offers a Wide Range of Improvements

Scientists in our laboratory have overcome the inherent limitations of symmetric PCR by developing an advanced form of asymmetric PCR, known as LATE-PCR.