Applied Biosystems QuantStudio 3D Quick Reference page 6

e. Allow the Digital PCR 20K Chip to sit on the heated
block for 20 seconds.
f. Add 10-15 drops of Immersion Fluid directly to the chip
so that the fluid covers the entire surface.
10-15x
7. Seal the Digital PCR 20K Chip:
a. Remove the red protective film from the QuantStudio
3D Digital PCR Chip Case Lid, then carefully place it
onto the prepared Digital PCR 20K Chip so that the
window in the lid aligns with the chip.
b. Pinch together all four sides and corners of the Chip
Case for at least 10 seconds to seal it. While pressing,
apply some lateral pressure to ensure a tight seal.
8. 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).
b. Using a lint-free laboratory wipe, remove any excess
Immersion Fluid from the Chip Case to ensure optimal
imaging on the QuantStudio
9. 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.
6
Correct Incorrect
Fluid over Fluid on Insufficient
chip edges fluid
>10 sec.
3D Instrument.
™
b. Power on the UV Stylus in the UV Stylus Stand by
twisting the cap near the power button.
c. Load the Digital PCR 20K Chip assembly into the UV
Stylus Stand, and allow the Chip Sealant to cure for
0.5-3 minutes. A longer exposure, no longer than
3 minutes, ensures a complete seal.
™
10. 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.
Thermal cycle the Digital PCR 20K Chips
Materials required
• Prepared QuantStudio
• ProFlex 2x Flat PCR System (or Dual Flat Block GeneAmp
™
PCR System 9700)
• QuantStudio
™
• QuantStudio
™
• QuantStudio
™
• Isopropanol
• Gloves, powder-free, nitrile
• Laboratory-grade, lint-free wipes
Perform the PCR
Note: Install the Tilt Base to the thermal cycler for the following
procedure.
1. Open the heated cover, then wipe the surface of both sample
blocks using a lint-free wipe to ensure that they are clean and
dry.
2. Confirm the Tilt Base is installed to the thermal cycler and
that Chip Adapters are installed to both sample blocks.
QuantStudio
3D Digital PCR 20K Chips
™
3D Digital PCR Chip Adapters
3D Digital PCR Thermal Pads
3D Digital PCR Tilt Base
3D Digital PCR System Quick Reference
™
>30 sec
UV
®