Chapter 5 Performing the Background Calibration and Optical Calibration
Troubleshooting
64 Applied Biosystems 7300/7500/7500 Fast Real-Time PCR System Installation and Maintenance Guide
Notes
Troubleshooting
Troubleshooting – Background Calibration
Condition: Cannot extract the data (background calibration failed)
Signals that exceed 72,000 fluorescent standard units (FSU) are considered beyond the limit of normal background
fluorescence for a 7300/7500/7500 Fast instrument. Such signals may indicate that either the background or the sample
block contains fluorescent contaminants. Common contaminants include: ink residue from permanent pens, powder from
disposable gloves, and dust.
To determine the source and location of the contamination:
1. In the plate document for the calibration:
a. Select the Results tab.
b. Select the Spectra tab.
2. Select all wells of the plate document.
3. Inspect the raw background data for an irregular spectral peak
or peaks.
Wells producing raw spectra that exceed 72,000 FSU are
considered irregular and could be contaminated.
a
b
Click to select
all wells
Possible
contamination
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