ABI Prism® 7900HT Sequence Detection System ... - OpenWetWare

Extracting Pure Dye
Information from
the Analyzed Run
7-20 System Maintenance
The purpose of viewing the data in the Pure Dye Wizard is to eliminate irregular pure
dye peaks from the data set. The wizard presents the spectral data from the pure dye
plate in sets of three wells, each containing the same pure dye. Because the wells
displayed by the wizard contain the pure dye at an identical concentration, the signal
peaks for the set should be identical. Occasionally, pipetting inaccuracies or
contamination can cause a well signal to shift slightly. While viewing the data, the
outlying peaks must be eliminated.
To extract the pure dye information from the run data:
Step Action
1 From the Analysis menu, select Extract Pure Dye Wizard.
The Extract Pure Dye Wizard dialog appears.
2 Follow the instructions as explained by the Extract Pure Dye Wizard to extract the
pure dye spectra.
When presented with each screen, do the following:
a. Inspect the spectra for shifts in peak location.
b. If the data set contains a outlying peak, eliminate it by clicking check box of the
associated well.
Note Dye spectra are generally acceptable if they peak at the same location as
their group but diverge slightly at other wavelengths.
c. Click Next to view the next three wells.
d. Repeat steps a to c for all remaining wells until prompted with a message
reporting the extraction of the pure dyes.
The software extracts the pure spectra and stores the data as a component of the
calibration file.
3 From the File menu, select Save.
The software saves the plate document.
4 From the File menu, select Close.
Wavelength
shift
Click here
to remove it
The software closes the plate document.
5 If performing spectral calibration of a 96-well block module, repeat the previous
procedures on pages 7-18, 7-19, and 7-20 to run the second Pure Dye plate.