Observation
Possible cause
Recommended action
Calibration failed but plate is
undamaged
The incorrect plate was used for
calibration performed.
Use the plate that matches the calibration
performed.
The plate was improperly
prepared.
Repeat the calibration with the plate properly
prepared.
If the calibration fails again, order a replacement
plate. Use Smart Help if the replacement plate
fails (see “Request technical support with Smart
Help” on page 159).
Identify contamination
Signals that exceed the limit of normal fluorescence may indicate fluorescent
contaminants on the calibration plate or the sample block. Common contaminants
include ink residue from permanent pens, powder from disposable gloves, and dust.
1.
View the calibration data and note the wells that failed the calibration.
2.
Remove the plate from the instrument, rotate the plate 180°, then perform the
calibration again.
3.
Determine the location of the failed wells again as in step 1.
Position of failed wells
Action
Identical
The sample block is contaminated.
Decontaminate the sample block (see page 146).
Reversed
The plate is contaminated.
Discard the plate, then perform the calibration using a
new calibration plate.
4.
Use Smart Help if the calibration fails after you decontaminate the sample block
and replace the plate (see “Request technical support with Smart Help” on
page 159).
Create a background plate
(optional)
Whenever possible, use a background plate listed in “Required materials to prepare
calibration plates” on page 84. These plates contain a buffer that accurately
simulates the reagents used for PCR, and, therefore, produces high-quality
calibration data.
If a background plate is not available, you can create one as described below.
Required materials:
• MicroAmp
™
optical 96-well reaction plate
• Optical adhesive cover or optical flat caps
• Pipettor, 200‐µL (with pipette tips)
• Powder‐free gloves
Chapter 7
Calibrate and verify instrument performance
Perform ROI/uniformity, background, and dye calibrations
7
90
QuantStudio
™
6 Pro and 7 Pro Real-Time PCR Systems User Guide
