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).
4.
In a 0.5- or 1.5-mL reaction tube, prepare sufficient PCR
reaction mix for your samples. Scale the component amounts
appropriately, depending on the number of samples that you
are running.
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 31.3 µL of PCR reaction mix to each labeled reaction
tube.
8.
Transfer 3.5 µL of each sample, diluted to the appropriate
concentration, to the corresponding 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
necessary, place the reactions on ice until you are ready to use
them. If placed on ice, warm the samples 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 Chips manually or
automatically using a QuantStudio
™
3D Digital PCR Chip Loader.
Load the chips using the Chip Loader
Materials required
• Prepared digital PCR reactions
• QuantStudio
™
3D Digital PCR 20K Chips
• QuantStudio
™
3D Digital PCR Chip Case Lids
• QuantStudio
™
3D Digital PCR Sample Loading Blades
• QuantStudio
™
12K Flex OpenArray
®
Immersion Fluid
• QuantStudio
™
12K Flex OpenArray
®
Immersion Fluid Tip
• QuantStudio
™
3D Digital PCR Chip Loader
• UV-Activated Chip Sealant Syringe
• Isopropanol
• Gloves, powder-free, nitrile
• Scissors
• Laboratory-grade, lint-free wipes
• Microcentrifuge
• Pipettes and tips, P10 to P1000
• Vortexer
Load and seal the chips using a Chip Loader
WARNING!
ULTRAVIOLET LIGHT HAZARD. Looking
directly at a UV light source can cause serious eye damage.
Never look directly at a UV light source and always
prevent others from UV exposure. Wear appropriate
protective eyewear and clothing.
IMPORTANT!
Wear powder-free gloves while preparing
QuantStudio
™
3D Digital PCR 20K Chips.
IMPORTANT!
The QuantStudio
™
3D Digital PCR Chip Loader
must be placed on a level surface to ensure consistent loading
across the Digital PCR 20K Chips.
1.
Prepare the workspace:
a.
Allow the prepared digital PCR reaction to equilibrate
to room temperature (approximately 15 minutes).
b.
Remove the following consumables from their
packaging and place them on a clean, dry, lint-free
surface:
• QuantStudio
™
3D Digital PCR Chip Case Lid
• QuantStudio
™
3D Digital PCR Sample Loading
Blade
2.
Prepare the syringe containing Chip Sealant:
a.
Remove the syringe, plunger, and tip from the
protective packaging.
IMPORTANT!
Do not discard the brown plastic bag
provided. When not in use, store the Chip Sealant
syringe within the bag and in a dark location.
b.
Remove the protective caps from both ends of the
syringe, twist and push the tip to lock it into place, then
insert the plunger into the opposite end of the syringe.
c.
Place the assembled syringe within its protective
package until ready for use.
IMPORTANT!
Confirm that the tip is locked firmly in place
before proceeding.
Keep out
of sunlight
3
2
1
4
1
Plunger
2
UV-Activated Chip Sealant Syringe
3
Chip Sealant Tip (twist to attach)
4
Brown plastic bag (do not discard)
2
QuantStudio
™
3D Digital PCR System Quick Reference