Thermo Scientific QuantStudio 1 - Page 63

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 gure 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.
Conrm that the liquid in each well is at the boom of the well and free of
bubbles. If it is not, centrifuge the plate again.
IMPORTANT! Keep the boom of the plate clean. Fluids and other contaminants
on the boom of the plate can contaminate the sample block and cause an
abnormally high background signal.
Chapter 5 Calibrate and verify instrument performance
Calibrate custom dyes
5
QuantStudio
™
1 Real-Time PCR System Installation, Use, and Maintenance Guide
63