Applied Biosystems QuantStudio 3D Quick Reference page 7

3. Load the QuantStudio
thermal cycler:
a. Place the sealed Digital PCR 20K Chips onto the sample
block so that the fill ports on the chips are positioned
toward the front of the thermal cycler.
IMPORTANT! If you are thermal cycling less than
24 Digital PCR 20K Chips, load according to the
following guidelines:
·
Load each Digital PCR 20K Chip so that the fill port
is oriented toward the front of the thermal cycler.
·
Load the right sample block first, placing at least
1 chip on the right sample block.
·
Balance the load between the left and right blocks so
that the pressure applied by the heated cover and
thermal pads is uniform across all of the loaded
chips.
b. Lay the QuantStudio
PCR 20K Chips.
IMPORTANT!
Make certain that each QuantStudio
Thermal Pad is centered on the sample block and that it
completely covers the Digital PCR 20K Chips.
2x
c. Slide the heated cover forward and pull the heated
cover lever down to engage the cover with the
QuantStudio 3D Digital PCR 20K Chips.
™
4. Use the thermal cycler to select and start the pre-
programmed run for the Digital PCR 20K Chips. See the
ProFlex PCR System User Guide (Pub. no. MAN0007697) or
™
the GeneAmp PCR System 9700 Base Module User Manual
®
(Pub. no. 4303481) for more information on running
methods.
Table 2 ProFlex 2x Flat PCR System PCR Method
™
PCR Protocol
Stage 1 Stage 2
96.0°C 60.0°C 98.0°C
0:10:00 0:02:00 0:00:30 0:02:00
1x 39x
QuantStudio 3D Digital PCR System Quick Reference
™
3D Digital PCR 20K Chips into the
™
Thermal Pads over the Digital
™
CAUTION HOT
Cover
Temp.
Stage 3
60.0°C 10.0°C
∞ 70.0°C
1x
Table 3 GeneAmp
Stage 1
96.0°C
10:00
1x
You can remove the Digital PCR 20K Chips from the block
immediately after the final extension step at 60°C is complete
and the temperature of the block is <30°C.
IMPORTANT!
hour after you remove them from the sample block. After
thermal cycling, the Digital PCR 20K Chips can remain on
the sample block for up to 24 hours.
Unload the thermal cycler
™
CAUTION!
operation, the sample block can be heated to 100°C. Before
performing the following procedure, be sure to wait until
the sample block reaches room temperature.
1. Raise the heated cover lever and slide the cover back to
expose the Digital PCR 20K Chips.
2. Remove the QuantStudio
block and set them on a clean, dry surface.
3. Remove the Chip Adapters from the thermal cycler sample
block and place them on a clean, dry surface. Remove the
Digital PCR 20K Chips from the Chip Adapters and allow
them to equilibrate to room temperature.
4. Visually inspect each Digital PCR 20K Chip for signs of leaks
or potential problems. Using a lint-free laboratory wipe,
remove any condensation or Immersion Fluid from the
surface of the chip by wiping in one direction. If necessary,
use a lint-free laboratory wipe sprayed with isopropanol to
remove any dried residue.
The Digital PCR 20K Chip is now ready to be imaged by the
QuantStudio 3D Instrument. Run the prepared Digital PCR 20K
™
Chips within 1 hour after removing them from the thermal cycler.
Rxn.
Analyze the chips
Vol.
Materials required
• Prepared Digital PCR 20K Chip
1 nL
• Isopropanol
• Gloves, powder-free, nitrile
• Laboratory-grade, lint-free wipes
PCR System 9700 PCR Method
®
PCR Protocol
Stage 2 Stage 3
60.0°C 98.0°C 60.0°C 10.0°C
2:00 0:30 2:00
39x 1x
Read the Digital PCR 20K Chips within one
PHYSICAL INJURY HAZARD. During
Thermal Pads from the sample
™
Run Rxn.
Speed Vol.
99:59 Std. 20 μL
7