Bio-Rad QX100™ Droplet Digital™ PCR System - Manual (Page 16)

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Chapter 2 Droplet Generation
QX100 Droplet Generator Instruction Manual
2.3 Preparation for PCR
1. Pipet 40 μl of the contents of the top wells (the droplets) into a single column of a 96-well plate.
Aspirating droplets from the DG8 cartridge. Dispensing droplets into a 96-well PCR plate.
Use the following pipetting techniques to avoid shearing or coalescing the droplets:
To aspirate droplets from the DG8 cartridge:
■ ■
Use an 8-channel manual P-50 Pipetman with 200 µl tips (not wide- or narrow-bore); do not use a P-20
or P-1000 Pipetman
■ ■
Place the cartridge holder on a flat surface and position the pipet tips in each of the 8 top wells at
a ~30–45° angle, vertical into the junction where the side wall meets the bottom of the well. Do not
position the pipet tip in a vertical orientation (90°) or against any flat surface of the well; do not allow the
tips to be flat against the bottoms of the wells
■ ■
Slowly draw 40 µl droplets into the pipet tip (should take ~5 sec, and ~5 µl air is expected); do not
aspirate >40 µl, as this causes air to percolate through the droplets
■ ■
Pipet slowly. Apply a stable resistive force to the plunger to draw and aspirate droplets smoothly into
and out of pipet tips
To dispense droplets into the 96-well plate, position the pipet tip along the side of the well — near, but
not at, the bottom of the well — and slowly dispense the droplets (~5 sec).
Cover the plate (for example, with 8-cap strips or the lid from a pipet tip box) as you work to prevent
evaporation and contamination with particulates.