DROPLET DIGITAL PCR (ddPCR)
Droplet digital PCR (ddPCR) is a refinement of the conventional PCR method. The ddPCR method is based on a water-oil emulsion droplet technology which enables a DNA sample to be divided into 20.000 nano-liter sized droplets. All of these droplets are now independent small PCR reactions each and the sum of all of these reactions are then used for the final result. It is the massive input of small reactions which is the key feature of the increase sensitivity achieved by this method. The method makes is possible to analyze each droplet as positive or negative based on fluorescence and can detect the presence of rare events down to 0.001% This method provides result with high precision with a less amount of sample and preform an absolute quantification of the specific target. This advanced technique is used to finding the needle in a haystack and is used for detection of Low frequency mosaic CNV ´s (Copy number variation), findings of epigenetic biomarkers for diseases diagnosis in blood, methylation specific detection of methylated sites, Non-invasive prenatal diagnosis of monogenic disorders and detection of somatic cancer mutations in blood samples.
ddPCR technique is developed by Bio-Rad.
Chemistry: ddPCRMultiplex Supermix, Bio-Rad.
Hardware: QX200 Droplet Digital PCR
Data processing: Data is analyzed in QX manager Software.