Thermo Scientific QuantStudio 1 - Calibrate Custom Dyes; Custom Dyes Overview; Use a Dilution Series to Determine an Optimal Custom Dye Concentration

Calibrate custom dyes
The instrument can run assays designed with custom dyes. Custom dyes include:
• Dyes that are not manufactured by Thermo Fisher Scientic.
• Dyes or formulations of dyes that are not system dyes for the instrument.
• For custom dye requirements, see “Custom dyes“ on page 10.
Workflow: Custom dye calibration
For each custom dye, determine the optimal dye concentration. Use this concentration
to prepare all subsequent dye calibration plates.
Use a dilution series to determine an optimal custom dye concentration
“Prepare a custom dye dilution plate“ on page 63
▼
“Run the dilution plate as an experiment“ on page 64
▼
“Determine the optimal dye concentration“ on page 64
▼
Calibrate the custom dye using the optimal concentration
Create a custom dye calibration plate
▼
Add a custom dye to the instrument
▼
“Perform a custom dye calibration“ on page 66
Custom dye dilution guidelines
Prepare a dilution series for each custom dye.
• Target several dye concentrations within a range of 100–2,000 nM.
• Choose a 2- or 3-fold dierence in dilution points.
• Dispense 10–20 µL per well in a 96-well plate.
• Dilute the dye in buer compatible with your master mix.
• (Intercalating dyes only) Add the appropriate amount of amplied PCR product to
generate uorescence.
Custom dyes
overview
Use a dilution
series to
determine an
optimal custom
dye concentration
Chapter 5 Calibrate and verify instrument performance
Calibrate custom dyes
5
62
QuantStudio
™
1 Real-Time PCR System Installation, Use, and Maintenance Guide