Agarose Jelly Lab by Katie Crecelius, grace henry, alana kaffenberger, ashtyn taylor

Procedure:

Pour an Agarose Gel

  1. In order to make a 1.2 % agarose gel, add 0.6 grams of agarose to 50 mL of the TAE buffer.
  2. Heat and stir the agarose solution on a hot plate using a magnetic stir bar for 2-4 minutes or until the solution is clear.
  3. Allow the agarose to cool for a minute so that it doesn't crack or warp the gel tray.
  4. Seal the ends of the gel-casting tray with either tape or dams and insert the well-forming comb.
  5. Pour 5 mm of agarose solution into the gel-casting tray and move bubbles to the side.
  6. Allow the gel to solidify completely (15-20 minutes).
  7. When it is solidified, place the gel in the electrophoresis chamber with the wells at the negative end.
  8. Fill the electrophoresis chamber with buffer until it completely covers the gel.

Loading the Gel

  1. Using a pipet, extract a sample of DNA from the vile and place it in a well of the agarose gel.
  2. Repeat the previous step with each DNA sample, using a separate pipet for each sample and placing them into a separate wells.

Running the Gel

  1. Close the top of the electrolytes chamber and connect the electrical leads to the power supply, connecting the red to the positive source and the black to the negative source.
  2. Turn on the power supply and set the voltage to 100 watts.
  3. Let the DNA sit until bands of the loading dye form, moving toward the electrophoresis apparatus.
  4. Allow the DNA to electrophorese until the blue band is about 1 cm from the end of the gel.
  5. Turn off the apparatus and move the gel to a staining tray.

Staining the Gel

  1. Cover the gel in the staining tray with CarolinaBLU Final stain and let sit for at least 20 minutes (but no longer than 45 minutes).
  2. Pour the stain back into the bottle.
  3. Rinse the gel with distilled water.
  4. Fill the tray with enough distilled water to submerge the gel.
  5. Continue this process with the distilled water to further distain the gel.
  6. Leave gels in the fridge overnight.

Results

Our bands did not properly form in the gel, so there are no results. Next time, the comb should be placed in the gel so that the wells do not go all the way through and can hold the DNA.

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