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PCR – A Powerful Tool for Multiple Applications

 

Microsynth offers a comprehensive workflow that begins at the sample extraction stage to produce high-quality data. Choosing to outsource your PCR application saves you time and effort, but also provides benefits from technical expertise and facilities dedicated to conducting various PCR-based experiments.

 

Digital PCR vs. Real-time PCR vs. end-point PCR at a glance:

  Digital PCR Real-Time PCR End-Point PCR
Overview Re-arrangement of the sample in thousands of nanoliter end-point PCR reactions allows determination of the absolute concentration of the analyte. Measures accumulation of the PCR amplification product as it occurs.

Measures the amount of accumulated PCR product at the end of the PCR cycles.
Quantitative? Yes, the compartmentalization of the analyte fits a Poisson distribution. Yes, data is collected during the exponential growth (log) phase of PCR when the quantity of the PCR product is directly proportional to the initial amount of analyte. No, though rough estimation of the amount of the accumulated PCR product gives 'semi-quantitative' results.
Applications
  • Absolute quantification of viral load
  • Absolute quantification of DNA or RNA
  • Rare allele detection (Mutation or SNPs in liquid biopsies)
  • Absolute quantification of gene expression
  • Genome Edit Detection
  • Copy number variation (i.e. quality control of CAR-T and other adoptive cell therapies)
  • Environmental monitoring
  • Residual DNA quantification
  • Quantitation of gene expression
  • Microarray verification
  • Quality control and assay validation
  • Pathogen detection
  • MicroRNA Analysis
  • Viral quantitation
  • siRNA/RNAi experiments

Amplification of DNA for:

  • Sequencing
  • Genotyping
  • Cloning
Summary

Digital PCR is the method of choice when:

  • references or standards are hardly available
  • you need a robust quantification (high tolerance to PCR inhibitors)
  • you aim to quantify rare targets in complex backgrounds
  • you aim to detect small fold changes, where real-time PCR fails

Real-time PCR is the method of choice when:

  • you measure the analyte in a very broad dynamic range up to 9 Logs
  • you expect analyte concentration change over 2-fold
  • you want to make cost-effective high-throughput screening

End-point PCR is the method of choice when:

  • you need accurate size-based discrimination
  • you amplify a specific locus of the DNA/RNA for more detailed analysis
  • you want to confirm presence of the specific locus in your DNA/RNA