Gene Expression and Proteomics

Gel Extraction

Protocol for Gel Extraction

1. Use a clean razor blade to cut DNA band. Place in microcentrifuge tube.
2. Add 1.4mL of Solution A from SpinPrep Gel DNA Kit into the microcentrifuge tube.
3. Incubate at 50°C for approximately 10 minutes until agarose melts. Be sure to mix by inverting periodically.
4. Cool down to room temperature by placing microcentrifuge tube at room temperature or in -20°C freezer for a few minutes.
5. Obtain a cartridge and receiver tube. Place cartridge inside the receiver tube. Load 700uL of DNA mixture into cartridge. Centrifuge. Remove solution from the receiver tube.
6. Load again, centrifuge, and remove solution. Repeat if necessary.
7. Add 350uL of Solution B and pulse to remove remaining ethanol (the remaining ethanol is an inhibitor for RE and ligase). Centrifuge for 2 minutes. Remove solution.
8. Place the cartridge in elute receiver tube. Add 40uL of Solution C (0.1x TE Buffer warm at 50°C). Incubate at 50°C for 2 minutes.
9. Centrifuge for 1 minute. Remove cartridge, label, and store.