Bio-Rad QX200™ AutoDG™ Droplet Digital™ PCR System - Manual (Page 13)

Instruction Manual | 5 4 | Automated Droplet Generator 4 | Automated Droplet Generator
1.4 System Setup and General Operation Instructions
3. Perform PCR — remove the 96-well PCR plate containing droplets from the Automated Droplet Generator,
seal the plate with foil, and perform PCR to end point (~40 cycles) using a PX1 ™ Plate Sealer and
C1000 Touch ™ Thermal Cycler (see Table 2).
4. Read droplets — load the plate into the QX200 (or 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 (–).
If reading or quantifying droplets and recovering material from droplets in parallel, prepare two sets of
reactions, one for each application. For example, a set of eight wells of droplets can be generated: four of
these will be read after thermal cycling, and four will not be read.
5. 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.
The QX200 ddPCR System is compatible with hydrolysis probe (TaqMan) chemistry and can detect up to
two fluorophores and up to four targets at a time (FAM/VIC or FAM/HEX). It is also compatible with EvaGreen
chemistry. Use only the approved Bio-Rad supermixes listed in Table 2 with this system; using unapproved
supermixes may harm the instrument and voids the warranty.
The QX100 ddPCR System does not support EvaGreen chemistry; running this chemistry will damage
the instrument.
1.4 System Setup and General Operation Instructions
Connect the Automated Droplet Generator to a power source using the power cord and power adapter
provided. Leave 2" (~5 cm) clear space behind and 5" (~13 cm) clear to the right and left of the instrument
for proper ventilation. Position the instrument such that it can be easily disconnected from the power
source, should that become necessary for servicing the equipment.
The Automated Droplet Generator is powered on by plugging it into a power source.