Prepare a custom dye dilution plate
IMPORTANT!
Wear powder-free gloves throughout the procedure.
1.
Prepare a 2- or 3-fold dilution series of the custom dye.
2.
Dispense aliquots of each dilution into the center of a reaction plate, then seal the plate.
A full plate is not needed.
See the following figure for suggested replicates.
100 nM
200 nM
400 nM
800 nM
1600
nM
3.
Vortex the plate for 5 seconds, then centrifuge at 750–1,000 ×
g
for 2 minutes.
4.
Confirm that the liquid in each well is at the bottom of the well and free of bubbles. If it is not,
centrifuge the plate again.
IMPORTANT!
Keep the bottom of the plate clean. Fluids and other contaminants on the bottom
of the plate can contaminate the sample block and cause an abnormally high background signal.
Run the dilution plate as an experiment
1.
Load the plate into the instrument.
2.
Set up a genotyping experiment on the instrument.
• Load an experiment created in the desktop software.
a. In the home screen, touch
Load Experiment
.
b. Select the experiment location, then the experiment file.
Chapter 5
Calibrate and verify instrument performance
Calibrate custom dyes
5
66
QuantStudio
™
1 Plus Real-Time PCR System Installation, Use, and Maintenance Guide
