User Manual

QX100 Droplet Reader and QuantaSoft Software Instruction Manual2 |
Chapter 1 QX100 Droplet Digital PCR System
1.2 System Components
The system consists of two instruments and associated software, consumables, and reagents (Table 1.1):
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QX100 droplet generator — utilizes proprietary reagents and microfluidics to partition samples into 20,000
nanoliter-sized droplets
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QX100 droplet reader — following PCR amplification of the nucleic acid target in the droplets, this
instrument analyzes each droplet individually using a two-color detection system (set to detect FAM and
either VIC or HEX); PCR-positive and PCR-negative droplets are counted to provide absolute quantification
of target DNA in digital form using QuantaSoft software
Additional materials required for performing ddPCR are listed in Table 1.2.
1.3 Droplet Digital PCR Workflow
Droplet Digital PCR involves the following steps (4.5–5.5 hours for the complete workflow):
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Prepare PCR-ready samples — combine nucleic acid sample (DNA or RNA), primers, and probes (FAM,
VIC, or HEX) with Bio-Rad ddPCR supermix (see Table 1.2)
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Make droplets — load 20 μl of the ddPCR reaction into the DG8
™
droplet generator cartridge, then load the
cartridge into the QX100 droplet generator to partition the sample into droplets. The QX100 droplet generator
uses microfluidics to combine oil and aqueous sample to generate the nanoliter-sized droplets required for
ddPCR analysis. It processes up to 8 samples at a time in under 2 minutes
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Perform PCR — pipet droplets from the cartridge to a 96-well PCR plate and seal the plate with foil. Perform
PCR to end point (~40 cycles) using a thermal cycler
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Read droplets — load the plate into the QX100 droplet reader and start your run. The droplet reader sips each
sample, singulates the droplets, and streams them in single file past a two-color detector. The detector reads the
droplets to determine which contain a target (+) and which do not (–)
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Analyze results — the droplet reader connects to a laptop computer running QuantaSoft software. The
software provides a complete set of tools for setting up and naming samples, running and controlling
the instrument, and analyzing results. It also reads the positive and negative droplets in each sample and
plots the fluorescence droplet by droplet. The fraction of positive droplets in a sample determines the
concentration of target in copies/μl
QX100 Droplet Digital PCR system. Left, QX100 droplet reader. Right, QX100 droplet generator.