Beckman Coulter CEQ 8000, User Manual

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Fragment Analysis Procedures
242 CEQ™ 8000 Genetic Analysis System
•
AE ver.1
is the calibration used for the CEQ Fragment Analysis test
sample.
•
SNP ver. 1
is used when performing a
SNP analysis
. The SNP analysis
will still proceed if the mobility correction is not selected, but the values
obtained may vary slightly.
•
No Correction
, which is the default dye mobility calibration, should be
used when no dye mobility calibration is desired. With this selection, the
system will not make any compensation for dye effects on fragment
mobility.
The CEQ Fragment Analysis test sample must be analyzed with the
AE ver.1 dye mobility calibration for correct sizing of the fragments.
2. Select the desired
Standard Mobility Reference
from the drop-down list box.
• The standard mobility reference is a calibration curve from an existing
analysis run under similar conditions.
• Use of a standard mobility reference provides information which will assist
in the identification of size standards when spurious peaks or other factors
make size standard identification problematic.
• Once a calibration curve has been generated and the coefficients
determined, it can be saved as a standard mobility reference to aid in the
identification of standard fragments in subsequent analyses of different
samples using the same standard.
If you have an existing set of results with good resolution, you may
want to save the parameters of the standard curve as a Standard
Mobility Reference to be used in subsequent analyses.
• In order for the curve to be saved as a standard mobility reference, the X
variable must be mobility.
• If the system fails to identify standard fragments during analysis, try
creating a new standard mobility reference, run under separation conditions
that more closely resemble those used in the failed analysis
Dye Spectra
Dye spectra values are calculations of the relative emission intensities of each dye in
each of the four detection channels. These values are calculated during each analysis,
and are used to transform the four electropherogram channels to dye signal traces for
each of the dye labels present in the sample.
When a dye spectrum cannot be calculated from the sample, the analysis process will
terminate unless system dye spectra are available.
The
Dye Spectra options
allow you to select an existing set of dye spectra to be
used instead of estimating the dye spectra from each sample.
This option can shorten the analysis time by up to 50%.