Beckman Coulter CEQ 8000, User Manual

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User’s Guide 235
• For calibrations that span a long size-range or extend into size regions where the
size selectivity of the separation system is very non-linear, a fourth degree
polynomial (
Quartic Curve
) may result in more accurate size estimates than a
cubic curve. This is usually the best choice for routine use with the
SizeStandard-600 set .
• The
Local Southern
method estimates the size of a sample fragment from its
mobility and the mobility's of the four standard fragments closest in size. If this
method is selected, the
X Variable
options are grayed-out and mobility is
automatically selected.
5. The Migration Variable fields are used to select either
migration time
or
mobility
as the independent variable for the calibration curve. Size is the
dependent variable.
• Selecting migration time will use the average migration times and sizes of
the standard fragments to construct a size calibration.
• Selecting mobility will use fragment velocity (cm/s) divided by the
separation potential (V/cm) with the sizes of the standard fragments to
construct a size calibration.
Fragment Analysis Parameters - Quantitation Tab
This tab allows you to set the parameters for calculating the quantity of all peaks by
comparing their heights or areas to a single peak of the same color whose quantity is
known.
Once you have entered the information for the known peak into this window, the
software will look for the reference peak in each trace using the specified size or time.
Based on this information, the software can then calculate the quantity of each of the
remaining peaks in the sample.
ID Standard
1. Within the
ID Standard
area of the window, use the option buttons to select the
method which the software will use to identify your reference peak.
• By selecting the
Size
option, the software will identify your reference
peak based upon the fragment size specified in the
Size
field listed for
each of the four dyes.
• By selecting the
Time
option, the software will identify your reference
peak based upon the migration time specified in the
Time
field listed for
each of the four dyes.