Bio-Rad Components for Older Model Spot Cutter - Manual (Page 13)

4.5 Making Cuts (Excisions)
Select the appropriate cutting tip action from the Operation menu, Others. A description
of the options is provided in Section 8.4.
Hint: For best deposit of gels and membrane spots, add 150–175 µl of liquid (water,
acetonitrile, etc.) to each microtiter plate well. This will provide the best condition to
deposit spots effectively, and it will keep the spots from drying out in the microtiter wells.
Recommendations for cutting tip actions:
Acrylamide Gels: Use standard (default) method. For increased assurance of positive eject,
select Double Eject.
PVDF Membrane: Use Eject With Turn in order to help eject the static membrane from
the stainless steel tip.
Calibration: Use Cut without Turn in order to keep the cutting tip from marking the
template.
A cutting run can be selected by either-
(i) Selecting the Run shortcut button located below the menu bar.
(ii) Selecting the Cutting menu, then selecting Run from the drop down menu.
You may not wish to cut all the spots selected during this cutting run. This can be achieved
by putting the desired starting spot number and last spot number in the "Start from" and "To"
fields of the Set-up Cut Spot window.
Note: An individual run must consist of a sequential subset of the total spots selected, i.e.
you can not select to cut spots 4 & 6 on an individual run without spot 5 being cut. An
individual cutting run cannot be out of order, ie. 4, 6, 5 is invalid, it would need to be 4, 5, 6.
Once the spot sequence to be cut has been selected, press the OK button of the Setup Cut
Spot window.
When cutting is complete, the Cutting Complete box will be displayed, select OK to
continue. It is advisable to check the completeness of the cutting run by visually verifying
that each well has a gel spot in it. If there are any missing wells, it is likely that the gel spot
is still in the gel. The cutter can very easily be directed to re-cut these spots. See next section
on Cut Previous Spot methods.
4.6 Re-Cut Previous Spot
If there is the need to re-cut a previously selected spot, there are several ways to direct the
spot cutter to re-cut a spot.
a. The MTP grid area of the screen can be used to easily re-select the few spots that may not
have been picked up from the gel. Move the dotted line box within the MTP grid to the
box containing the spot to be deleted by using the arrow keys on the keyboard.(You may
need to right click on the table to produce the dotted line bos. Hit escape, then move the
box with the arrow keys.) Right click on the MTP spot destination window located at the
bottom right-hand corner of the screen. From the pop-up menu select Single Cut. The
cutting arm will go to that spot location, perform the cut, and return to the appropriate
MTP well location. This can be repeated for other individual spots to be re-cut.
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