User guide

In this procedure, you will prepare the PCR reactions for the samples that you intend
to load to QuantStudio
®
3D Digital PCR 20K Chip
. The volumes of the example
protocol have been adjusted so that two chips are run for each sample.
Table 3 PCR Reaction Mix
Material Stock conc.
Volume (μL)
Final conc.
Per chip
1 sample/
2 chips
[1]
QuantStudio
®
3D Digital PCR
Master Mix
2X 7.25 17.4 1X
TaqMan
®
Assay (primer/probe
mix)
20X 0.725 1.74 1X
Diluted DNA 10 ng/μL 5.0
[2]
12.0 3.45 ng/μL
Water — 1.525 3.66 —
Total volume — 14.5 34.8 —
[1]
V
olumes include 20% excess to compensate for volume loss from pipetting.
[2]
The recommended DNA volume is based upon a human gDNA sample at 10 ng/µL concentration with the
target sequence present at two copies per diploid genome. This recommended volume will vary depending
upon species, sample type, and sample concentration. For best results, we recommend adding sufficient DNA
such that the concentration of target sequence in the final reaction is between 200 and 2,000 copies/µL.
1.
Remove the following from storage and allow them to warm at room
temperature:
• QuantStudio
®
3D Digital PCR Master Mix
• TaqMan
®
Assay(s)
2.
Review the concentration of your DNA samples, and prepare dilutions if
necessary.
3.
Gently invert the tube of Digital PCR Master Mix 10 times (or gently vortex on
low-medium speed).
Prepare the
reaction mix and
samples
Chapter 2 Prepare Samples and Load Reactions
Prepare the digital PCR reactions
2
QuantStudio
®
3D Digital PCR System User Guide 39