Bio-Rad EXQuest Spot Cutter - Manual (Page 60)

7.0 LANE/VOLUME EXCISION TOOL
The Lane/Volume Excision Tool is designed to provide a simple and efficient method for cutting lanes and
bands from 1D gels and blots. This tool is available only with the purchase of a full Quantity One
® software
license.
7.1 SETTING UP A CUT RUN
1. Make sure the spot cutter is set up as described in Sections 2 and 3 and that the required cutting
head is connected to the spot cutter.
2. Prepare the gel/blot as described in Section 4.
3. Open the Quantity One software and click Excision / Lane/Volume Excision Tool….
Figure 7-1. Excision pull-down menu
4. Make sure that the DI water bottle is full and the waste bottle is empty at the start of the run and
click Prime Pump.
5. If cutting blots, fill three 1.5 ml capless microtubes with water to approximately 1 mm below the tube
top and then place the tubes in the three membrane tip wash wells. After making a cut, the cutting
tip will be dipped into one of the three tubes after each cut to wash the tip and remove any
membrane plugs that have adhered to the tip.
6. In the "Acquire Image" panel (see Section 8.1), select White Trans for visible stained gels such as
Coomassie or silver stain, or UV EPI for fluorescent stains such as SYPRO Ruby.
7. Close the spot cutter front door and click Acquire Image to take an image of the gel tray contents.
Do not open the door during image acquisition. As a safety precaution, the UV EPI lights will
automatically turn off if the door is opened. Images acquired with "UV EPI" are automatically
inverted by the software to display dark spots with a light background. If the image does not detect
enough spots or is too faint, use the transform function to optimize the image contrast (see Section
8.3).
8. Identify the cutting tip size, 1.0 mm or 1.5 mm, in the "Cut Options" panel (see Section 8.4). Note
that the cut request circle size displayed on the image depends on the tip size selected.
9. From the "Cut Options" panel, specify the type of gel to cut (see Section 8.4).
• Free gels. Select this option to use a protocol optimized for cutting non-backed 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.
• 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.
LANE/VOLUME EXCISION TOOL
7-1