Agarose Jelly Lab by Katie Crecelius, grace henry, alana kaffenberger, ashtyn taylor
Pour an Agarose Gel
- In order to make a 1.2 % agarose gel, add 0.6 grams of agarose to 50 mL of the TAE buffer.
- 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.
- Allow the agarose to cool for a minute so that it doesn't crack or warp the gel tray.
- Seal the ends of the gel-casting tray with either tape or dams and insert the well-forming comb.
- Pour 5 mm of agarose solution into the gel-casting tray and move bubbles to the side.
- Allow the gel to solidify completely (15-20 minutes).
- When it is solidified, place the gel in the electrophoresis chamber with the wells at the negative end.
- Fill the electrophoresis chamber with buffer until it completely covers the gel.
Loading the Gel
- Using a pipet, extract a sample of DNA from the vile and place it in a well of the agarose gel.
- 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
- 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.
- Turn on the power supply and set the voltage to 100 watts.
- Let the DNA sit until bands of the loading dye form, moving toward the electrophoresis apparatus.
- Allow the DNA to electrophorese until the blue band is about 1 cm from the end of the gel.
- Turn off the apparatus and move the gel to a staining tray.
Staining the Gel
- 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).
- Pour the stain back into the bottle.
- Rinse the gel with distilled water.
- Fill the tray with enough distilled water to submerge the gel.
- Continue this process with the distilled water to further distain the gel.
- Leave gels in the fridge overnight.