How often should a spectral calibration be run on the Applied Biosystems 3730 Series DNA Analyzer?
There is no fixed time as to when a spectral must be run. However, some users prefer to run a new spectral at a preset time to ensure it is up to date. Data quality will be the largest indicator of when a new spectral is required. A separate spectral calibration needs to be performed for each dye set, array type, and array length. For G5-RCT dye set users only: a spectral calibration must be re-run if the array detection cell is moved/repositioned. Refer to FAQ "When should I run a spectral calibration?" for more information on when a spectral should be run.
Find additional tips, troubleshooting help, and resources within our Capillary Electrophoresis Instruments Support Center.
When should I run a spectral calibration on the Applied Biosystems 3730 Series DNA Analyzer?
A new spectral calibration should be run:
- Whenever a new dye set is used on an instrument.
- After any optics adjustment performed by a service engineer.
- If the instrument has been moved.
- If you see a decrease in spectral separation (pull-up and/or pull-down peaks).
- If you alter any conditions (dye set, array type, or array length).
- For G5-RCT, Any4dye-HDR, and S dye set users only: Any time the array detection cell is moved/repositioned.
Find additional tips, troubleshooting help, and resources within our Capillary Electrophoresis Instruments Support Center.
What is a spectral calibration?
A spectral calibration is an algorithm applied to raw data, which converts it into the component 4 or 5 dye data stored in the sample files. A spectral is created for a specific dye set (combination of dyes), array type (4 or 16 capillaries), and array length (36cm or 50cm). It is used to correct for the natural overlap of the fluorescent dyes.
Find additional tips, troubleshooting help, and resources within our Capillary Electrophoresis Instruments Support Center.
Which fragment analysis matrix standards can I use for my Applied Biosystems 3130 Series instrument?
DS-02 (Dye Set E5), DS-30 (Dye Set D), DS-31 (Dye Set D), DS-32 (Dye Set F), and DS-33 (Dye Set G5) are all supported on the Applied Biosystems 3130 Series systems. Please refer to the Applied Biosystems 3130/3130xl Genetic Analyzers Getting Started Guide (http://tools.thermofisher.com/content/sfs/manuals/cms_041468.pdf) for more information.
Find additional tips, troubleshooting help, and resources within our Capillary Electrophoresis Applications Support Center.
I am using only one dye for fragment analysis but I see peaks in other colors below my peak of interest. Why is this?
If the peaks in other colors are directly below the peak of interest, the issue could be that the fluorescent dye being used is not part of the selected dye set, the spectral calibration needs to be performed, or the peaks are offscale. Confirm that the dye set selected on the instrument is compatible with the dye being used, run a new spectral calibration if the correct dye set has been selected and, if the signal intensity is too high, decrease sample concentration during PCR or when preparing samples for electrophoresis.
Find additional tips, troubleshooting help, and resources within our Capillary Electrophoresis Applications Support Center.
