ThermoFisher Scientific QuantStudio 1, Инструкция по установке, эксплуатации и обслуживанию

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Calibrate custom dyes
Custom dyes overview
The instrument can run assays designed with custom dyes. Custom dyes include:
• Dyes that are not manufactured by Thermo Fisher Scientific.
• 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 66
▼
“Run the dilution plate as an experiment” on page 66
▼
“Determine the optimal dye concentration” on page 67
▼
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 69
Use a dilution series to determine an optimal custom dye concentration
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 difference in dilution points.
• Dispense 10–20 µL per well in a 96
‑
well plate.
• Dilute the dye in buffer compatible with your master mix.
•
(Intercalating dyes only) Add the appropriate amount of amplified PCR product to generate
fluorescence.
Chapter 5
Calibrate and verify instrument performance
Calibrate custom dyes
5
QuantStudio ™ 1 Plus Real-Time PCR System Installation, Use, and Maintenance Guide
65