Objective 

The objective of this week’s lab was to make an agarose gel and put our eDNA through the purification process.

Purpose

The purpose of making an agarose gel was so we could “run” it. Running the gel allows us to see the bands of DNA from our samples under a UV light. The purpose of purifying our DNA is to ensure that we are only running DNA and not any environmental impurities.

Procedure

1. Obtain 4 mL of 10x TAE with a serological pipette and transfer into a 50 mL conical tube
2. Fill the conical tube up to 40 mL with DI water
3. Weigh out 0.4f grams of agar powder and transfer from the balance into a 125-mL Erlenmeyer flask
4. Pour the solution from the conical tube into the Erlenmeyer flask and microwave for 1 minute
5. After microwaving, swirl flask for about 1 minute to ensure that all agar powder is dissolved in the 1x TAE and DI water solution
6. Have the TA add 2 microliters of Ethidium Bromide to the Erlenmeyer flask and swirl flask a little to mix everything
7. Pour the solution in the Erlenmeyer flask into a premade gel mold
8. Wait about 30 minutes for the gel to solidify
9. Once the gel is solidified, pour a thin layer of 1x TAE over the gel to ensure it does not dry out and harder
10.  Place the labeled mold in a plastic bag and store in refrigerator until next lab

Cleanup and Storage

The conical tube was rinsed and left on the prep area to dry and the Erlenmeyer flask was bleached, rinsed, and set on the drying rack. The gel molds were placed in the refrigerator to keep for the next week. All trash was picked up and area was wiped down.

Future Goals

My main future goal is to be able to successfully use the process of gel electrophoresis to see the bands of DNA from our samples. I was told these hardly ever work so it would be really cool if we got ours to be successful.

Other Notes

DNA purification was performed by lab partners Madison Ambrose and Megan Hudson