When To Perform A Spectral Calibration - Thermo Scientific appliedbiosystems SeqStudio Flex Series User Manual

When to perform a spectral calibration

Perform a spectral calibration if you do any of the following actions.
• Use a dye set/polymer combination that you have not previously calibrated
• Use a custom dye set (a dye set that is not available from Thermo Fisher Scientific) or a dye set
definition that is not factory-provided
• Observe a decrease in spectral separation (pull-up/pull-down in peaks) in the raw or analyzed data
• After performance of maintenance by a service representative that affects the optical systems
• After the instrument is moved
• You frequently run samples for which auto-spectral calibration cannot be performed (these samples
generate a "Calibration from manual..." calibration status)
Under most conditions, manual spectral calibrations are not needed after the capillary array is changed
because of the auto-spectral calibration feature (see "Auto-spectral calibration during an injection"
page 150). However, in applications with overlapping peaks that are labeled with different dyes, such as
microsatellite instability, the spectral characteristics may not meet the requirements of the auto-spectral
calibration algorithm. If an auto-spectral calibration cannot be performed and no saved auto-spectral
calibration is available for the capillary position, the most recent manual spectral calibration is applied to
the samples. When routinely running samples for which auto-spectral calibration cannot be performed,
it is important to perform a manual spectral calibration after you change the capillary array or move the
instrument, to provide a valid manual calibration for samples.
Note: For sequencing applications, you can skip the spectral calibration process, and perform the
sequencing install run. If you select Keep Spectral Calibration Data in the install run, the software runs
a spectral calibration for the dye set during a sequencing check and saves the updated dye matrix as a
manual spectral calibration. For information, see "Perform an install run"
calibrations are not required for sequencing applications.
SeqStudio ™ Flex Series Genetic Analyzer with Instrument Software v1.0 User Guide
Chapter 12 Run calibrations and install checks
Run a spectral calibration
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12
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328. Routine manual
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