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LANE/VOLUME EXCISION TOOL 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/tubes as this will allow cut membrane to float between adjacent wells. 10. Select the vessel type that will be used from the "Plates Options" panel (see Section 8.5) and specify the well volume and loading order. The well volume will determine the maximum number of spots that can be placed in a single well/tube. 11. Optional. Check Hydrate in the "Hydration Options" panel to automatically hydrate the gel during the cut run. Specify how often it is to be hydrated (see Section 8.6), then click Define hydration area to identify the gel or blot location. Gels and membranes should typically be hydrated at least once every hour. 12. If gels are being cut, enter the wash volume required to clean the gel cutting tip (50 to 500 µl, see Section 8.7) in the "Wash Options" panel. 7.2 CUTTING LANES Entire Lane mode is used to define the lanes from which cuts will be made. 1. Select Entire lane in the "Specify Cuts" panel. Figure 7-2. “Specify Cuts” panel, “Entire lane” selected 2. 7-2 Click Frame Lanes and then click in the image to create lanes in an adjustable frame. In the pop-up box that appears, enter the number of lanes to create. Check or uncheck Show excisions on image to show or hide cut request labels on the image