Load and seal the Digital PCR 20K Chips
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. Follow the
manufacturer’s recommendations for appropriate
protective eyewear and clothing.
IMPORTANT!
Wear powder-free gloves while preparing the
QuantStudio
™
3D Digital PCR 20K Chips.
1.
Prepare the workspace:
a.
Preheat the heated block to 40°C, the optimal
temperature for loading Digital PCR 20K Chips.
b.
Allow the prepared digital PCR reaction to equilibrate
to room temperature (approximately 15 minutes).
c.
If necessary, replace the batteries of the UV Stylus in the
UV Stylus Stand.
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, push and twist 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 package until
ready for use.
IMPORTANT!
Confirm that the tip is locked firmly in place
before proceeding.
3.
Prepare a syringe containing QuantStudio
™
12K Flex
OpenArray
®
Immersion Fluid:
a.
Remove the syringe, plunger, and tip from the
packaging.
b.
Before uncapping the syringe, gently pull back the
plunger 1-2 mm and release it to break any resistance
that may have formed during storage.
c.
Uncap the syringe, then attach the OpenArray
®
Immersion Fluid Tip by pushing it into place.
d.
Carefully depress the plunger until Immersion Fluid
flows from the tip of the assembled syringe, then place
it on a clean surface until ready for use.
IMPORTANT!
Use all of the Immersion Fluid within
60 minutes of uncapping the syringe. Once a syringe is
opened, you cannot reattach the cap for later use.
4.
Remove the Chip Case Lids and Sample Loading Blades
from their packaging and place them on a clean, dry surface.
5.
Open the QuantStudio
™
3D Digital PCR 20K Chip package,
gently grasp the chip by its sides, then place it face-up on the
heated block, preheated to 40°C.
IMPORTANT!
Allow the Digital PCR 20K Chip to sit on the
heated block for at least 5 seconds before loading it.
40±1°C
6.
Load the PCR reaction onto the Digital PCR 20K Chip:
a.
Briefly vortex and centrifuge the prepared reaction
(master mix, assay, and template), then aspirate 14.5 µL
of the solution into a P20 pipette. If the reactions were
placed on ice, allow them to warm prior to loading.
b.
While holding the Sample Loading Blade by its edges,
carefully dispense the reaction into the sample-loading
port. After loading, gently tap the top of the Sample
Loading Blade to spread the reaction across the full
length of the blade.
IMPORTANT!
Do not depress the pipette to the second
stop. Doing so increases the chance of introducing an
air bubble to the Sample Loading Blade.
c.
While holding the Sample Loading Blade at a 70-80°
angle relative to the heated block so that the port faces
up, place the edge of the blade at the end of the Digital
PCR 20K Chip. Adjust the angle of the blade until you
visually confirm that it is wetting the chip. Then, in one
smooth motion, slowly drag the blade across the chip
while applying a slight downward pressure to dispense
the reaction.
IMPORTANT!
Move the Sample Loading Blade slowly.
The time required to move the blade across the chip
should be approximately 10-12 seconds.
40±1°C
70-80°
10 sec.
d.
Confirm that the Sample Loading Blade is empty.
QuantStudio
™
3D Digital PCR System Quick Reference
5