An Efficient PCR Protocol for co-Amplification of Multiple Markers

Dor Yuval , HUJI, School of Medicine - IMRIC, Developmental Biology and Cancer Research


Multiplex PCR, PCR, cfDNA, Research tool, Liquid Biopsy, DNA methylation, cell-free DNA

Current development stage

TRL5 Technology validated in relevant environement


PCR on bisulfite-treated DNA, followed by next generation sequencing, has a major potential as a diagnostic tool, by identifying DNA derived from specific tissue based on cell type-specific methylation markers. To maximize assay sensitivity, multiple loci have to be amplified in parallel. However, multiplex PCR of bisulfite-converted DNA remains a major challenge.

Our Innovation

A new protocol that allows to efficiently co-amplify a large number of loci after bisulfite conversion (multiplex PCR), to generate material that is ready for sequencing without further manipulation.

  • simultaneously amplifying over 30 different and independent methylation markers
  • Increased specificity and sensitivity: detects low levels of strongly diluted differentially methylated loci
  • Compatibility with smaller amount of available bisulfite-converted template
  • Reduced costs and time of procedure


  • Most important tool for simultaneous detection of a large number of markers of the same tissue/state, increasing sensitivity and specificity of the test while reducing overall reagent and labor costs
  • Method validated in multiple studies, including reactions using genomic DNA and second bisulfite-converted cfDNA - the most challenging template
  • Allows for routine, affordable non-invasive detection of tissue specific cell death in the human body

Contact for more information:

Mel Larrosa
VP Business Development Healthcare
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