Gel Preparation:

  1. Weigh 1.0 g agarose in a weigh boat
  2. Tare a 250 mL beaker and transfer the agarose to the beaker
  3. Add 50 mL of 0.5x TBE-Buffer to the 250 mL beaker
  4. Place the beaker in the microwave and heat for 1-2 minutes until dissolved
  5. Refill the beaker with ddH20 to 50 g
  6. Add 400 uL of 1.375M MgCl2 and 2 uL of ethidium bromide to the beaker. Swirl the contents carefully while holding the beaker
  7. Pour the gel contents into the gel rig and add the comb. Remove the comb after about 20 minutes. Turn the gel so that the wells are on the left side.
  8. Pour 250 mL of .5x TBE-Buffer with magnesium into the gel rig. The wells are now ready to be loaded

Gel Preparation

Gel Loading:

  1. Add 3 uL of loading dye to every 15 uL of sample. Add 3 uL of loading dye to 1.5 uL of scaffold and 13.5 uL of ddH20 to create a reference scaffold
  2. Pipet 6 uL of ladder into the first well, 17 uL of reference scaffold into the second well, and 17 uL of sample into the following wells
  3. Connect the anode on the side with the wells and cathode from the rig to the power source and add ice around the gel rig to prevent overheating
  4. Run the gel at 70mV for 2 hours

Gel Loading

Gel Imaging

  1. Transfer the gel from the rig to the UV table
  2. Place the camera on the UV table so that it covers the gel and turn on the camera
  3. Use the imaging program on the computer to take a centered, clear picture of the gel
  4. Adjust the camera resolution and brightness as needed

Gel Imaging