Thermo Scientific appliedbiosystems SeqStudio Flex Series - When to Perform a Spectral Calibration

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 pr
eviously 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 aects 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” on
page 150). However, in applications with overlapping peaks that are labeled with dierent 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” on page 328. Routine manual
calibrations are not required for sequencing applications.
Chapter 12 Run calibrations and install checks
Run a spectral calibration
12
SeqStudio
™
Flex Series Genetic Analyzer with Instrument Software v1.0 User Guide
311