Protocols
Gel Electrophoresis
Pouring a Gel:
- Determine gel size and well size, insert cassette into tray
- Add 0.01% SYBR-Safe to the final gel volume (about 30mL gel volume for a 10-well gel → 1–3uL SybrSafe)
- Mix with the pipette tip, moving bubbles to a corner
- Cover to cool for 10–15 min
- Check to make sure the gel is fully solidified and appears uniform before removing the well comb
- Turn the gel cassette so the wells are oriented toward the negative electrode (run to red)
- Cover with 1X TAE until the gel is fully covered by a few mm of buffer
Running a Gel:
- Plug in cassette, run at 100V
- Check to make sure you are running to RED
- Check to make sure bubbles are forming at electrodes
- Cover to run
- Run for 15–20 minutes, but check loading dye progress
Visualizing a Gel:
- Remove the gel from the electrophoresis chamber
- Place in the viewing machine, select blue/UV light and set time to ~1 second
- Adjust exposure until just a few pixels are saturated
- Take picture and label the gels
- Turn off the blue/UV light and clean everything
