Bio-Rad EXQuest Spot Cutter User Manual
Page 47
a.
From the "Automatically Select Gels to Cut" screen, set the Tolerance. The tolerance value is
used to determine the spot quantity range that is acceptable when choosing the gel from which
a particular spot will be cut. If the quantity of a spot in only one gel falls within the tolerance, that
gel will be auto-selected for the spot cut. However, if the quantity of a spot in two or more gels
falls within the tolerance range, then the auto-select mechanism will consider which gels will
best minimize the time and effort required to switch gels on the spot cutter platform. For
example, if you have five gels in an analysis set with the tolerance ratio set to 1 (the lowest
setting), auto-selection specifies different cuts in all five gels, requiring you to change gels in the
spot cutter five times. However, by increasing the tolerance ratio slightly, auto-select finds that
three gels contain spots whose quantities fall within the specified tolerance, and it adjusts the
cuts so that only those three gels are selected. This will save you considerable time and effort
switching gels in the spot cutter. As you adjust the tolerance, monitor the number of gels
selected by the tolerance setting.
b Check
the
Include quality box to consider spot quality when auto-selecting spots. When this
checkbox is selected, a well-resolved spot of sufficient quantity will be favored over a less
well-resolved spot of higher quantity (within the specified tolerance ratio). After checking the
Include quality box, look to see if it has changed the number of gels selected from which cuts
will be made.
6.
Click Next to Specify the Excision Options.
Figure 6-8. “Specify Excision Options” Screen
7.
Identify the cutting tip size, 1.0 mm or 1.5 mm, in the "Cut Options" panel (see Section 8.4).
8.
Specify the type of Material to cut (see Section 8.4) in the "Cut Options" panel,
•
Free Gels. Select this option to use a protocol optimized for cutting unbacked gels.
•
Plastic-backed Gels. Select this option to use a protocol optimized for cutting plastic-backed
gels. When this option is used, gel plugs are cut and then dislodged from the gel backing by a
side-to-side cutting tip motion.
•
Glass-backed gels. Select this option to use a protocol optimized for cutting glass-backed gels.
When this option is used, gel plugs are cut and then dislodged from the gel backing by a
side-to-side cutting tip motion. Note that the cutting tip is raised by 3 mm to accommodate the
glass plate.
ANALYSIS SET EXCISION TOOL
6-5