User manual

18 | Droplet Digital
™
PCR Applications Guide
Fig. 2.4. Loaded DG8 cartridge.
Fig. 2.5. Loaded DG8 cartridge placed in the QX200 droplet generator.
Note: Each DG8 cartridge generates eight wells of droplets. Any unused wells on the
cartridge must be filled with 1x ddPCR buffer control.
The Bio-Rad ddPCR supermixes have been formulated specifically to work with the droplet
chemistry. Altering the components used in the QX100 droplet generator or using a different
supermix will negatively impact results. A 1x final concentration of supermix must be used
for proper droplet formation and proper target quantification.
After loading a 20 µl PCR reaction, load 70 µl of droplet generation oil into the bottom wells
of the DG8 cartridge (Figure 2.4). Attach a gasket across the top of the DG8 cartridge and
place it into the QX200 droplet generator (Figure 2.5). The droplet generator produces
about 20,000 droplets per sample in about 2.5 min for eight samples. Droplets should
be transferred to a 96-well PCR plate by pipetting gently.
We recommend designing your experimental plate layout on a 96-well plate in columns
because the cartridge contains eight wells.
PCR
After generating droplets in the DG8 cartridge, pipet the droplets from the top wells of the
cartridge into a PCR plate. The PCR plate should be heat sealed using Bio-Rad’s PX1
™
PCR plate sealer and pierceable foil heat seal.
Note: Using an alternative seal with glue can damage the droplet reader.
Designing Droplet Digital
™
PCR Experiments