d.
Dispense the fluid so that it
sweeps in and around the fill
reservoir toward the vent port.
When pipetting the reagents into
the array card, pipet the entire
100-µL volume into the fill
reservoir, but do not go past the
first stop of pipettor plunger or
you may blow the solution out of
the port.
IMPORTANT!
Do not allow the tip to contact and possibly damage the
coated foil beneath the fill port.
4.
Centrifuge and seal the array card as explained in “Fill the calibration TaqMan
Create a custom dye plate for calibration
The QuantStudio
™
6 or 7 Flex System can be used to run assays designed with custom
dyes (dyes not manufactured by Thermo Fisher Scientific). Custom dyes must excite
between 455 and 672 nm and read between 505 and 723 nm.
Before using custom dyes with the QuantStudio
™
6 or 7 Flex System, you must:
• Determine optimum dye concentration.
• Create a custom dye plate.
• Add the custom dye to the software.
• Perform a dye calibration.
• Centrifuge with plate adapter
• Custom dye(s)
• Safety glasses
• Powder-free gloves
• MicroAmp
™
Optical 96/384-Well Reaction Plate
• Optical Adhesive Cover
• Pipettors and pipette tips (200-µL and 1000-µL)
• Tubes (2-mL and 10-mL)
• Deionized water
Note:
Wear powder-free gloves while creating the dye plate.
1.
Prepare and load the custom dye plate:
a.
In the center of a 96- or 384-well plate, prepare a dilution series of the
custom dye (for example, 25, 50, 100, 200, 400, 800, 1600, and 3200 nM) using
20 µL volumes for a 96- or 384-well plate.
b.
Seal the reaction plate using an optical adhesive cover.
Before you use
custom dyes
Required
materials
Determine
optimum dye
concentration
Appendix F
Calibration consumable preparation
Create a custom dye plate for calibration
F
QuantStudio
™
6 and 7 Flex Real-Time PCR Systems v1.6 Maintenance and Administration Guide
153
