Auto-Spectral Calibration Requirements - Thermo Scientific appliedbiosystems SeqStudio Flex Series User Manual

changes in the spectral characteristics from the original manual calibration dye matrix. The instrument
performs an auto-spectral calibration for system and custom dyes.
The auto-spectral calibration process is iterative. For all capillaries in each injection, the instrument
evaluates the spectral characteristics of the current sample.
• If the spectral characteristics of the current sample meet the requirements for the auto-spectral
calibration algorithm—The instrument optimizes, updates, saves, and applies the auto-spectral
calibration to the current sample data.
• If the spectral characteristics do not meet the requirements for the algorithm—The instrument
applies the auto-spectral calibration from a previous injection to the current sample data. If an
auto‑spectral calibration is not available for the capillary position, the instrument applies the manual
spectral calibration to the current sample data.

Auto-spectral calibration requirements

For successful auto-spectral calibration, each dye used in the sample must contain at least one peak
that does not overlap with peaks that are labeled with a different dye.
In applications such as microsatellite instability, high peak density and overlap across dyes and a low
number of scans in the region of usable data prevents successful auto-calibration.
In other applications with high peak density, such as human identification, cell line authentication, or
sequencing, auto-spectral calibration is typically successful because of minimal overlap across dyes.
In samples where auto-spectral calibration cannot be performed and no saved auto-spectral calibration
is available for the capillary position, the instrument applies the most recent manual calibration to
samples. Over time, you may observe pull-up peaks for samples that cannot use auto-spectral
calibration. To address this issue, perform a manual spectral calibration if you frequently observe
pull-up peaks in sample, or after you change the capillary array or move the instrument.
SeqStudio ™ Flex Series Genetic Analyzer with Instrument Software v1.0 User Guide
Chapter 6 Start and monitor a run
Auto-spectral calibration during an injection
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