Applied Biosystems QuantStudio 3D Quick Reference page 4

5. Rotate the Chip Loader arm so that the Chip Case Lid solidly
contacts the Digital PCR 20K Chip, firmly press down for
15 seconds to ensure a tight seal, then press the Lid Nest
button and return the Chip Loader arm to its original
position.
IMPORTANT!
While applying the lid:
·
Press down on the Chip Loader until you reach a hard
stop. The arm requires >20 lbs of force to apply the Chip
Case Lid. If you suspect that you are applying insufficient
pressure, increase the duration of the press (>15 seconds).
·
Press the Lid Nest button before you retract the Chip
Loader arm.
·
Release the Chip Loader arm gently and do not press
down again to avoid creating a vacuum.
6. Fill the Chip Case with Immersion Fluid:
a. Hold the Chip Case by its edges and at a 45° angle so
that air can escape from the loading port as you fill it,
then carefully dispense Immersion Fluid into the port
until the Chip Case contains an air bubble no larger
than the fill port (<2-3 mm in diameter).
After filling the Immersion Fluid, rotate it slightly to
reveal any air bubbles that might be hidden behind the
serial number label. If bubbles are present, add
additional Immersion Fluid to remove them.
b. Using a lint-free laboratory wipe, remove any excess
Immersion Fluid from the Chip Case to ensure optimal
imaging on the QuantStudio
7. Seal the Chip Case using Chip Sealant:
a. Insert the syringe tip into the fill port of the sealed Chip
Case, then carefully fill the port with Chip Sealant,
ensuring that the fluid touches the walls of the hole.
When full, finish the seal by lifting the syringe tip and
add a small amount of sealant over the top of the port.
4
>15 sec
3D Instrument.
™
b. Insert the Digital PCR 20K Chip assembly into the UV-
Curing Station on the Chip Loader. Push the chip into
the station until the ultraviolet light illuminates.
IMPORTANT!
Do not allow the Digital PCR 20K Chip
to contact the roof of the UV-Curing Station.
c. When the light powers off (approximately ³15 seconds),
remove the chip and place it on a clean, dry, lint-free
surface.
8. Visually inspect the sealed Digital PCR 20K Chip for leaks,
bubbles, and correct lid orientation. Store the prepared
Digital PCR 20K Chip in a clean, dry, dark location until you
are ready to load it onto the thermal cycler.
Thermal cycle the prepared Digital PCR 20K Chips within 2 hours
after loading them.
IMPORTANT!
Store the prepared Digital PCR 20K Chips in a
clean, dry, dark location until you are ready to load them onto the
thermal cycler.
Load chips manually
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
™
• QuantStudio 12K Flex OpenArray
™
• UV-Activated Chip Sealant Syringe
• UV-Curing Stylus for Chip Sealant
• UV-Curing Stylus Stand
• Isopropanol
• Gloves, powder-free, nitrile
• Heated block (capable of maintaining 40±1°C)
• Scissors
• Laboratory-grade, lint-free wipes
• Microcentrifuge
• Pipettes and tips, P10 to P1000
• Vortexer
QuantStudio
UV
Immersion Fluid
®
Immersion Fluid Tip
®
3D Digital PCR System Quick Reference
™
>15 sec