Process of ddPCR
Set up ddPCR reaction mix with primers or probes, sample, Bio-Rad supermix (probes or EvaGreen). More information about setting up reactions.
Load sample plate on Bio-Rad Automated Droplet Generator. 20,000 droplets are generated in each well distributing target and background DNA into droplets and placed in a new plate. Oil type corresponds to either Probes or EvaGreen chemistry.
Seal plate immediately with foil in plate sealer to prevent evaporation of oil.
Run sample reactions on PCR using a slow ramp time of no more than 2-2.5°C/second.
Load PCR plate in QX200 Droplet reader. All droplets are read and counted. Analyze concentrations with QuantaSoft™ software. A copy of the software for use in your lab can be requested from CQLS staff.
There are three different ways the instrument can be run:
Full Service - Core Reaction plus Run: Investigator drops off samples along with primers and probes. CQLS staff set up the reactions and run them on the ADG, PCR and droplet reader
Partial Service – Core Run: Investigator sets up reactions and hands plate to CQLS staff to run on the ADG, our PCR and droplet reader with CSV file of information related to samples. Time of run must be coordinated with Staff and instrument availability.
Multi-User Service - Multiuser Run: Investigator reserves time to run system, sets up reactions, receives enough consumables to run ADG with their samples (oil, DG32™ Automated Droplet Generator Cartridges, specific tips for ADG, plate, foil), sets up droplets on ADG, seals plate, runs PCR on their own thermal cycler, reads droplets on QX200 Digital Droplet Reader. User must be trained by CQLS staff before using system.