Droplet digital PCR provides absolute quantification of target DNA or RNA molecules without the use of standard curves and can be run with EvaGreen or hydrolysis probe-based chemistries.
The BioRad QX200 combines water-oil emulsion droplet technology with microfluidics. The AutoDG droplet generator partitions each sample into 20,000 uniform nanoliter-sized droplets and a droplet reader bundled QuantaSoft™ software count the PCR-positive and PCR-negative droplets. ddPCR could be used to:
- Small fold difference, enable to measure 1.2x difference.
- Rare targets, low gene expression, low abundance microRNA and single cell analysis
- Absolute quantification
- Strong inhibition in samples.
Following image demonstrates the main steps in ddPCR.