dd PCR is an advanced version of PCR that provides simultaneous clonal amplification and fluorescence-based quantification of nucleic acids. A typical reaction in ddPCRTM is partitioned into several thousands of water-in-oil droplets where individual reactions are carried out providing absolute, precise and reliable quantification. The system includes: i) QX 200TM Droplet Generator; ii) T100TM Thermal Cycler; iii) PX1TM PCR Plate Sealer and iv) QX 200TM Droplet Reader. The reaction utilizes nucleic acids, primer sets, fluorescent probes (TaqMan probes with FAM and HEX or VIC), and proprietary supermix for droplet generation. The QX 200TM Droplet Generator generates approximately 20,000 nanoliter-sized uniform droplets which are transferred to a 96-well plate for clonal amplification. After the reaction, the droplets are read in the QuantasoftTM software bundled in QX 200TM Droplet Reader that counts PCR-negative and PCR-positive droplets and provides an initial DNA concentration of target molecule by statistical analysis in terms of copies/µL. ddPCRTM has demonstrated usage in the detection of rare DNA targets, determination of copy number variations and measurement of gene expression levels.
