Applied Biosystems 3500 - Section 6.1 Run spatial and spectral calibrations

Section 6.1 Run spatial and spectral calibrations
Run a spatial calibration
The software uses images collected during the spatial calibration to establish a
relationship between the signal emied by each capillary and the position where that
signal falls on, and is detected by, the CCD camera.
Perform a spatial calibration after you:
• Remove or replace the capillary array.
• Replace the capillary when it expires.
Note: When the instrument reads the information from a newly installed
capillary array, you are required to run a spatial calibration and a spectral
calibration before you can run plates.
• Open the detector door or move the detection cell.
• Move the instrument.
IMPORTANT! Do not open the instrument door during a spatial calibration run.
Doing so will stop the run and require you to restart the software.
1.
Preheat the ov
en if you will be selecting the Fill option for the calibration (ll the
array with polymer).
2.
Select Calibration4Spatial.
Note: The screen does not display results unless you have previously performed
a spatial calibration.
3.
Select No Fill, or select Fill to ll the array with polymer before starting the
calibration.
4.
Select Perform QC Checks.
5.
Click Start Calibration.
During the calibration, the software performs quality checks and calculates the
following values:
Attribute Calculation Threshold
Average peak height (sum of all peak heights)
divided by
(number of peaks)
• 8-cap: 6,400 RFU
•
24-cap: 3,000 RFU
Individual peak height Peak height 1,000 RFU
Uniformity (peak height
similarity)
(standard deviation)
divided by
(average peak height)
0.2
Capillary spacing max. spacing - min. spacing 2 pixels
Spatial calibration
overview
When to perform a
spatial calibration
Perform a spatial
calibration
Chapter 6 Run calibrations and install checks
Run a spatial calibration
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3500/3500xL Genetic Analyzer User Guide—Data Collection Software v3.1