c.
Load the prepared reaction plate.
2.
Start the calibration wizard:
a.
In the Home screen of the QuantStudio
™
Real-Time PCR Softw
are, click
Instrument Console.
b.
In the Instrument Console, select your system, then click Add to My
Instruments.
c.
Select the system, then click Manage Instrument.
d.
In the Instrument Manager, click Maintenance, then click ROI.
e.
In the ROI Calibration screen, click Start Calibration.
f.
In the ROI dialog box, click Next until prompted to load the system. When
the side door opens, load the sealed plate. Ensure that the plate or the array
card is properly aligned in the holder.
g.
In the ROI dialog box, select Check the box when the ROI calibration plate
has been loaded, click Next twice, then click START RUN to start the
calibration.
3.
When the run is complete, inspect the ROI images:
a.
Select the rst lter from the Filter drop-down list.
b.
Record the coordinate of the well that contains the lowest concentration of
dye and that is encircled by a ring. This well contains the optimal
concentration of the custom dye at the given lter.
c.
Repeat steps a and b for the remaining lters.
d.
After you determine the optimum concentration for each lter, determine
the optimum concentration for the custom dye. Compare the results from all
lters, then select the concentration that yields the highest possible signal in
all lters.
4.
Discard the plate.
WARNING! PHYSICAL INJUR
Y HAZARD. During instrument
operation, the plate can reach 100°C. Allow the plate to reach room
temperature before removing.
5.
In the ROI dialog box, click Finish to complete the calibration, then click No
when prompted to sav
e the results.
Appendix F Calibr
ation consumable preparation
Create a custom dye plate for calibration
F
154
QuantStudio
™
6 and 7 Flex Real-Time PCR Systems v1.6 Maintenance and Administration Guide
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