PCR reactions, in 20 μl volume, consisted of 9 μl cfDNA, 0.3 μM of each primer, 0.1 μM of each probe, and 2× ddPCR Supermix (Bio-Rad). The mixture was loaded into sample wells of an eight-channel disposable droplet generator cartridge (Bio-Rad). An additional 70 μl of droplet generation oil (Bio-Rad) was loaded into the oil well for each channel. After droplet generation the cartridge was removed and manually transferred with a 200 μl pipet to a 96-well PCR plate. The plate was heat-sealed, placed on a thermocycler (Bio-Rad T100) with the following program: 95 °C 10 min, (94 °C 30 sec, 56/58/60*°C 1 min) × 45 cycles (2°C/sec ramp rate each step during the cycling), 98 °C 10 min, 10 °C forever. After PCR, the 96-well PCR plate was read on the QX-100 droplet reader (Bio-Rad). Analysis of the ddPCR data was performed with QuantaSoft analysis software (Bio-Rad) that accompanied the droplet reader.
Eight channel disposable droplet generator cartridge
Manufactured by Bio-Rad
The eight-channel disposable droplet generator cartridge is a lab equipment product designed for use in droplet-based applications. It provides a platform for generating discrete droplets in a microfluidic system.
Automatically generated - may contain errors
Lab products found in correlation
Quantasoft analysis software, by Bio-Rad (3 mentions)
Droplet generation oil, by Bio-Rad (3 mentions)
Ddpcr master mix, by Bio-Rad (2 mentions)
Droplet generator, by Bio-Rad (2 mentions)
Droplet reader, by Bio-Rad (2 mentions)
Ddpcr supermix, by Bio-Rad (1 mentions)
Qx100 droplet reader, by Bio-Rad (1 mentions)
3 protocols using eight channel disposable droplet generator cartridge
1
Droplet Digital PCR for cfDNA Analysis
2
Digital Droplet PCR Protocol
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For performing the digital droplet-PCR (ddPCR) we prepared the reaction mixture by combining a 2x ddPCR Mastermix (Bio-Rad), 20x primer, and probes (final concentrations of 900 and 250 nM, respectively; Integrated Data Technologies) and template in a final volume of 20 μl (Supplementary Table 3 ). Then, each ddPCR reaction mixture was loaded into the sample well of an eight-channel disposable droplet generator cartridge (Bio-Rad). A volume of 60 μl of droplet generation oil (Bio-Rad) was loaded into the oil well for each channel. The cartridge was placed into the droplet generator (Bio-Rad), and the droplets were collected and transferred to a 96-well PCR plate. The plate was heat-sealed with a foil seal, placed on the thermal cycler and amplified to the endpoint (40 cycles). After PCR, the 96-well PCR plate was loaded on the droplet reader (Bio-Rad). Analysis of the ddPCR data was performed with QuantaSoft analysis software (Bio-Rad).
3
Digital Droplet PCR Protocol
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