Bio-Rad EXQuest Spot Cutter User Manual

Page 59

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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.

PVDF: This option is available only when the spot cutter is in membrane cutting mode, and is
used to specify that either a PVDF or nitrocellulose membrane will be cut.

Tips for cutting membranes

Fill microplate wells with DI water to within 1 to 2 mm of the top. In order to release the cut
membrane into a microplate well, the cutting tip must be in contact with water.

Do not overfill the wells or tubes, as this will allow cut membrane to float between adjacent
wells.

15. Optional. Select Make multiple cuts on large objects in the "Cut Options" panel (see Section 8.4)

to harvest large amounts of protein from a single spot and deposit it into a single well or tube. Enter
the maximum number of cuts that you want to allow in the Maximum cuts/object field.

16. Select the microplate format to be used, from the "Plate Options" panel (see Section 8.5), and then

specify the well volume and well loading order. The well volume will determine the maximum number
of spots that can be placed in a single well/tube.

17. Review the "Run Info" panel (see Section 8.8) for the number of cuts selected, the number of plates

required and the required plate well volume.

18. Press Proceed to finish creating the cut request list and to open the "Plates for Cut Run" dialog (see

Section 8.9). Enter the plate name and barcode/plate ID in their respective fields. The plate barcode
can be entered using a barcode scanner or typed in. To reserve wells on a microplate, click the Edit
button for the desired plate and then click on individual wells to reserve them.

19. Finish setting up the run as described in Section 6.2.

ANALYSIS SET EXCISION TOOL

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