Digital PCR Applications:
- Copy number variation
Copy number variation (CNV) analysis measures the number of sequence repeats for a specific genomic location with respect to a normal or reference genome. qPCR is an established tool for standard copy number variation analysis, with sufficient resolution for variants with 0 to 5 genomic copies. However, digital PCR provides more sensitivity when quantifying small fold copy number changes and discriminating between complex somatic copy number variations.
- Rare mutation detection
Rare mutation detection aims to identify a very low fraction of mutant targets in abundant wild-type targets. Detection of rare mutations can be particularly challenging because probes for mutants usually only differ from wild type by a single base. Digital PCR overcomes the challenge of probe-sequence similarities by digitizing into micro-reactions, thereby enriching for the targets. Also, dPCR is 100 times more sensitive than conventional methods for this type of analysis, with the ability to pool samples for even higher sensitivity.
- Liquid biopsy
Monitoring biomarkers via liquid biopsy is a promising tool in clinical research. In clinical cancer research liquid biopsy is used to measure circulating tumor DNA (ctDNA) as an indicator of therapeutic response, residual tumor burden, and potential therapeutic resistance. With its high precision and sensitivity for quantitative detection of rare targets, digital PCR can effectively discriminate ctDNA among high levels of background DNA in liquid biopsy samples.
- Cell and gene therapy research
Measure common viral titer backbones, custom genes of interest, or the presence of residual DNA without the use of a standard curve for faster, simpler, more precise quantification.
Vector copy number—assess the number of integrated vector genomes during characterization studies and monitor product stability and identity |
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Virus quantification—precisely quantitate genomic titer, viral particle titer, and full/empty capsid ratios during purification and lot release to accurately asses dose |
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Purity & safety testing—identify product and process-related impurities such as residual DNA or microbial contamination for in-process and release testing
Accurately measure viral load and pathogens or other biomarkers in environmental samples such as wastewater by absolute quantification with improved performance compared to qPCR in the face of common PCR inhibitors.
Digital PCR provides precise quantification of next-generation sequencing libraries to help maximize data quality and output. Because no other samples or standard curves need to be run, digital PCR removes a source of variability to help ensure accurate results.
Detect genome editing events in a fast and reliable way, especially those created using nucleases that cause double-stranded breaks, such as CRISPR-Cas9. Precise quantification of these nucleic acid editing events is crucial to determine the activity and efficiency of these enzymes in cells. |
dPCR, liquid biopsy, gene expression, genotyping, microRNA, mutation detection, gene therapy, copy number, MAP16