• Always wear powder-free gloves when handling Chip Cases.
IMPORTANT! Never handle the Chip Cases without gloves. Oils from your hands can contaminate the Digital PCR 20K Chips and interfere with imaging. • Handle the loaded Digital PCR 20K Chips by gently gripping the sides of each
Chip Case, being careful to not apply pressure to the Digital PCR 20K Chip within it.
• Do not touch the surface of the QuantStudio® 3D Digital PCR Chip Case Lid.
If you accidentally touch the surface of a Chip Case Lid, clean the Chip Case using a laboratory wipe that has been thoroughly sprayed with isopropanol, then dry it by wiping in one direction using a clean low-lint wipe.
Note: Debris or excess sample on the Chip Case Lid can interfere with thermal uniformity during thermal cycling, or fluoresce during imaging.
• Thermal cycle loaded Digital PCR 20K Chips within 2 hours after loading them.
X0300036
Note: Install the Tilt Base to the thermal cycler for the following procedure.
CAUTION! PHYSICAL INJURY HAZARD. During 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.
IMPORTANT! Thermal cycle prepared QuantStudio® 3D Digital PCR 20K Chips
within 2 hours after loading them.
IMPORTANT! Thermal cycling of Digital PCR 20K Chips must be performed on a ProFlex™ 2x Flat PCR System or a Dual Flat Block GeneAmp® PCR System 9700 as
described in this guide. No other thermal cyclers can be substituted at the time of the release of this manual.
1. Open the heated cover, then wipe the surface of both sample blocks using a low- lint 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.
IMPORTANT! You must install Chip Adapters to both sample blocks when thermal cycling Digital PCR 20K Chips, even if you are using only one block.
Guidelines for
handling Chip
Cases
Perform the PCR
Thermal cycle the Digital PCR 20K Chips
3. Load the QuantStudio® 3D Digital PCR 20K Chips into the 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. The fill port must be elevated during thermal cycling to prevent leaks from improperly sealed chips.
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.X0300036
Front of instrument CAUTION HOT 24x
·
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® Thermal Pads over the Digital 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.
IMPORTANT! Do not allow the QuantStudio® Thermal Pads to become
dirty. Fluids, debris, and other contaminants that adhere to the pad can transfer to the Digital PCR 20K Chips during thermal cycling and fluoresce during imaging.
CAUTION HOT
2x
c. Close and engage the heated cover with the QuantStudio® 3D Digital PCR
20K Chips.
IMPORTANT! Verify the heater cover is completely engaged to ensure a proper seal.
Chapter 3 Perform the PCR
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.
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 <25°C.
IMPORTANT! Read the Digital PCR 20K Chips within one 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 at 10°C.
Note: If you programmed your thermal cycler to perform a 10°C hold following thermal cycling, do the following to prevent condensation:
1. Confirm the final extension step at 60°C is complete, then stop the run:
·
ProFlex™ System – Press Stop Run, then press OK.·
GeneAmp® PCR System 9700 – Press STOP twice , then press Exit.2. Allow the thermal cycler to sit for at least 10 minutes with the heated lid closed.
3. Remove the chips.
Use a low-lint wipe sprayed with isopropanol to remove any remaining condensation or immersion fluid from the lids before imaging them.
CAUTION! PHYSICAL INJURY HAZARD. During 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. Open the heated cover to expose the Digital PCR 20K Chips.
2. Remove the QuantStudio® Thermal Pads from the sample block and set them on
a clean, dry surface.
IMPORTANT! Do not discard the QuantStudio® Thermal Pads. They can be
reused multiple times.