User guide

4.
In a 0.5- or 1.5-mL low-binding reaction tube, prepare sufficient PCR reaction
mix for your samples. Prepare the reaction mix at room temperature and scale
the component amounts appropriately, depending on the number of samples
that you are running.
Note: The following example assumes that you will load each sample to two
Digital PCR 20K Chips.
Material
Volume (μL)
1 sample/
2 chips
[1]
10 samples/
20 chips
[1]
QuantStudio
®
3D Digital PCR Master Mix, 2X 17.4 174.0
TaqMan
®
Assay(s), 20X (primer/probe mix) 1.74 17.4
Water 3.66 36.6
Total volume 22.8 228.0
[1]
V
olumes include 20% excess to compensate for volume loss from pipetting.
5.
Using a permanent marker, label a 0.5- or 1.5-mL reaction tube for each sample
that you intend to run.
6.
Vortex, then briefly centrifuge the DNA samples.
7.
Transfer 22.8 μL of PCR reaction mix to each labeled reaction tube.
8.
Transfer 12 μL of each sample, diluted to the appropriate concentration, to the
corresponding low-binding reaction tube. Mix well by gently pipetting up and
down after each transfer (or gently vortex on low-medium speed).
9.
Cap the reaction tubes, then briefly centrifuge them and immediately proceed to
load the Digital PCR 20K Chips.
IMPORTANT! For optimal results, load the Digital PCR 20K Chips as soon as possible
after setting up the reactions. If you placed the reactions on ice, warm them to room
temperature prior to loading.
Load the Digital PCR 20K Chips
After preparing your PCR reactions, you can load them into QuantStudio
®
3D Digital
PCR 20K Chip
s manually or automatically using a QuantStudio
®
3D Digital PCR Chip
Loader.
To load and seal Digital PCR 20K Chips:
• Automatically — See “Load the chips using the Chip Loader “ on page 42.
• Manually — See “Load chips manually“ on page 50.
Chapter 2 Prepare Samples and Load Reactions
Load the Digital PCR 20K Chips
2
40 QuantStudio
®
3D Digital PCR System User Guide