Protocol for Capturing an Image of an Agarose Gel Stained with
SYBR Safe using the UVP GelDoc-it PC based system.
Introduction
The gel imaging system used in the Nephrology Research Laboratory is the UVP
GelDoc-it PC based System with VisionWorks™LS Acquisition and Analysis
Software. The system has a light-tight darkroom with a full access safety door,
including a UV protective gel viewport and epi-illumination white light. The UV
transilluminator is housed inside the darkroom cabinet. The filter tray contains
filters for SYBR Safe, SYBR green, SYBR gold and ethidium bromide. For safety
reasons SYBR Safe DNA gel stain is the only gel stain currently permitted for use
in the lab. The image is taken using a 10 bit high resolution CCD camera and
printed using a Sony thermal printer.
Materials
Labcoat, Safety glasses and nitrile gloves
Agarose gel with SYBR Safe DNA gel stain to be imaged
Metal carrying tray
Tissue paper from white roll
UVP GelDoc-it System
Sony thermal printer
Printer paper (Part Number xxxxx)
Safety notes
 Read and sign the COSHH risk assessment, task risk assessment and
SOP for ‘Gel Electrophoresis using SYBR Safe’, before beginning the
procedure.
 The Laboratory rules must be adhered to at all times.
 Fastened up lab coat, safety goggles and disposable nitrile glove must be
worn.
 Do not open lab doors or use computers or telephones with gloves on.
 CAUTION! – The ultraviolet transilluminator is a powerful source of
UV radiation that will cause serious damage to unprotected eyes and
skin. Always make sure the darkroom door is firmly shut when the
transilluminator is switched on.
 Electrical hazard! Never use electrical equipment with wet gloves on.
Keep water and buffer away from all electrical connections. Do not use an
electrical connection if it is wet.
 Report all spills and faults with the equipment to the Safety Officer or your
line manager.
 Dispose of all waste according to the laboratory Disposal Policy.
Procedure for capturing a gel image using the UVP GelDoc-it System
1. Using the gloved hand, place the gel to be imaged onto the
transilluminator as described in the protocol ‘Electrophoresis of a SYBR
Safe gel’. Using the ungloved hand, ensure that the safety door of the
darkroom cabinet is securely shut.
2. Using the ungloved hand, turn on the computer and log in.
3. Double click on the Vision works icon to open it up.
4. Enter the Experimental data that you wish to record.
5. Open up the file.
6. Turn on the darkroom using the switch on the right hand side. Allow 30
seconds for the darkroom to initialise.
7. Click the icons on the top tool bar to open up the darkroom and the control
panel. Select ‘SYBR Safe Gel’ from the top drop down box. The SYBR
Safe filter is selected and the transilluminator switches on automatically.
8. Open the viewport window on the darkroom door to observe the gel. Close
the window.
9. Return to the PC and observe the image on the screen.
10. The image may be adjusted by opening or closing the aperture, or by
increasing or decreasing the exposure time. Adjustments to the image are
quite slow - Allow 10 seconds for any adjustments to be applied to the
screen. The image may be zoomed and focused if required.
11. Select ‘Stop/Capture’ to capture the image.
12. Go to the icon on the top menu bar ‘new area of intrest’. Use the mouse to
draw a box around the image by clicking and holding at the top left corner
of the gel image and dragging to the bottom right corner. Release the
mouse.
13. Go to ‘File’ → ‘Duplicate/Crop’. The cropped image is displayed. This may
be enhanced by adjusting the contrast etc. if desired.
14. Once the image is ready to print, click on the print icon, check that the
Sony printer is selected and click ‘print’.
15. Remove the printed image by tearing from the printer.
16. Save the gel image into your folder if necessary.
17. Switch off the darkroom using the switch on the right hand side of the
cabinet.
18. Close down the software down and turn off the computer.
19. Still using the ungloved hand open the darkroom door. Using the gloved
hand, remove the gel from the transilluminator into the metal tray lined
with tissue paper. Wrap the gel in the tissue and place into the yellow
clinical waste bin for disposal. Wipe the surface of the transilluminator with
tissue paper dampened with MilliQ water. Wipe dry with fresh tissue.
20. Using the ungloved hand close the darkroom door securely.
21. Remove glove and wash hands thoroughly at the hand washing sink
before leaving the lab.