Protocol for
Cutting PCR Bands out of an Acrylamide Gel
1. Run product out on acrylamide gel. !!!Remember to Rain-X the short plate!!!
2. Separate plates and stain gel with agarose overlay (if using unlabeled primers).
3. Mark the long plate using a green sharpie. (In order to line up the plate after scanning)
4. In the Scanner Control window for Flora, click on Setup. Select the orientation labeled “cutout orientation” below.
5. Remove overlay (if using one) and scan gel.
6. In ImageQuaNT, open the gel file. Under the View Menu, select “Actual Size”. Adjust the contrast and line the gel up on the screen to include your area of interest.
7. Under the File Menu, select “Print”. Make sure that the print menu is set to “Magnification” and “100%”, and hit “OK”.
8. Line up the printed gel image under the plate using the green sharpie marks.
9. Cut out bands using a spatula or scapel. Put each fragment into a 1.5 mL eppendorf with 200 uL TLE. Remember to clean spatula between each fragment.
10. Mix tubes by inverting or vortexing and leave o/n @ room temp.