Bio-Rad iQ™5 Optical System Software, Version 2.1 - Manual (Page 121)

Section 7 Calibrating the Instrument
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The following reagents and accessories are required to calibrate the iQ5 instrument:
• iCycler iQ calibrator dye solution kit (catalog #170-8792), which contains iQ5-specific pure
dye solutions and external well factor solution
• 3 x 96-well PCR plates or preferred reaction vessel
• Optical-quality sealing tape or preferred sealing method
NOTE: Calibrator dye solutions are NOT required for calibrating the MyiQ system.
7.3 Preparing Calibration Plates
Preparation of an External Well Factor Plate
Preparation of this plate is required for mask alignment and persistent well factor generation for
the MyiQ2, MyiQ and iQ5 systems.
To prepare an external well factor plate:
1. Dilute the 10x External Well Factor Solution to 1x with water.
2. Pipet 25 µl of the 1x external well factor solution into each of 96 wells of your preferred
reaction vessel.
3. Seal using your preferred sealing method you will use in your future experiments.
Preparation of a Background Calibration Plate
Preparation of this plate is required for background calibrations for the MyiQ2, MyiQ and iQ5
systems.
Leaving the wells completely empty, seal 96 wells of your preferred reaction vessel with your
preferred sealing method.
Preparation of the Pure Dye Calibration Plate
Preparation of this plate is required for pure dye calibration for the MyiQ2 and iQ5 systems.
To prepare a pure dye calibration plate:
1. In clean wells, pipet 25 µl of the appropriate 1x calibrator pure dye solution into 8 wells of
your preferred reaction vessel.
2. Repeat for all fluorophores you will use in your future experiments. See example of Pure
Dye Plate Setup.
3. Seal using your preferred sealing method you will use in your future experiments.
NOTE: Pure Dye Calibration is NOT performed on the MyiQ Real-Time PCR Detection System.
7.4 Performing Calibrations
The MyiQ2, MyiQ and iQ5 systems are calibrated using the calibration plates described in section
7.3 above and using the Calibration Module of the iQ5 software. Allow the instrument optics to
warm up for 20 min prior to performing the mask alignment, background calibration, persistent
well factor collection, or pure dye calibration procedures.