Appendix G/F12

From OpenWetWare
Jump to navigationJump to search
Wellesley College

BISC110/112- Introduction to Cell Biology Lab- Fall 2012


APPENDIX G. Instructions for Capturing Digital Images of Nucleic Acid Gels Stained with SYBR Safe Using the BioRad Imaging System

Instructions for Taking a DNA gel image stained with Sybr Safe using the BioRad ChemiDoc MP System with Image Lab Software

IMPORTANT: Ethidium Bromide stained gels may NOT be imaged in this equipment. Remove gloves and wash hands BEFORE using the computer. DO NOT contaminate the computer. The XcitaBLue Conversion Screen should be kept covering the UV transilluminator and only removed TEMPORARILY when using fluorphores other than Sybr Safe or SybrGreen. Please return the Xcita Blue Screen to the UV transillumintor if you remove it.

Quick and Easy Protocol for photographing Sybr Safe DNA gels:
1)Make sure the Power Button on the right front of the imager shows a green light. If not press it until the green light comes on and wait 5-10 min for warm up.

2) Open the UV transilluminator drawer on the lower front of the imager and make sure that the XcitaBlue Conversion Screen is in place. If not find it and position it covering the glass. Clean it with water and a paper towel if necessary.

3) Position your gel in the center of the glass.

4) Close the drawer, remove gloves, and wash your hands before using the computer to the left of the imager.

5) Open the ImageLab 4.0.1 software by double clicking on the icon on the computer desktop

6) Find and open the Protocol called SybrSafe under Recent Protocols or from the Open Menu. Double click to open it.

7) Click Position Gel (yellow button) to check the position of your gel.

8) Click Run Protocol (green button) to take the photo.

9) To Save your image (if you are not going to analyze it quantitatively), find or make a folder for your work in Documents (NOT on the Desktop). go to File---Export---Export for Publication (use the defaults, e.g. 300dpi). You will see a pop up message reminding you that this option is not optimized for analysis---it doesn’t matter—Click OK. Check the Location where your image will be saved, the FILE Name, and use the drop down menu to SAVE AS TYPE tiff or jpg. Click Save.

10) Close the Image Lab Software 11) Don gloves, remove your gel, clean the glass with water and a papertowel. 12) The computer AND the ChemiDoc Imager should remain ON. DO NOT Turn OFF the power or shut down or log off the computer.