Purpose/Objective

The purpose of today’s lab was to prepare DNA for amplification and to begin setting up our agarose gel for gel electrophoresis. The lab introduced us to putting together all of the primers, water, DNA, and buffers into a single solution to prepare to DNA Amplification.

Procedure

Preparing DNA Tubes

1. Obtain 3 tubes which are pre-filled with 12.5 μL of 2X Master Mix. Label the tubes so that you know which tubes will serve as your positive and negative controls, and which will serve  as your sampled DNA tube.
2. Add 1 μL of COX1  primers,  and 11.5 μL of water to the negative control tube.
3. Add 1 μL of COX1 primers, 5 μL of Paramecium DNA, and 6.5 μL of water to the positive control tube.
4. Add 1 μL of COX1 primers, 5 μL of sample DNA, and 6.5 μL of water to the sample DNA tube.
5. Store the tubes away for next week.

Preparing Agarose Gel

1. Measure out 0.6 grams of Agarose powder on a scale. Empty the powder into an Erlenmeyer flask.
2. Measure out 40 mL of T.A.E. and empty into the Erlenmeyer flask. Swirl the flask to thoroughly mix the solution.
3. Cover the top of the Erlenmeyer flask with weighing paper and a loose-fitting cap, and microwave the solution for 1 minute and 20 seconds at 70% power.
4. Remove carefully, and allow to cool in a water bath at 55 degrees Celsius for 5 to 6 minutes.
5. Add 2 μL of ethidium bromide to the solution. This will cause the solution to bind to the DNA.
6. Pour the solution into a gel rack on a flat surface to cool and solidify into gel.
7. Clean lab area and say bye to Will.

Conclusion

This lab was a pretty productive lab that was also fun. We didnt’ have any observable data for this lab, as all we did was prepare our DNA tubes and agar gel for next lab. Next week I believe that we will pour our DNA onto the gel, and run a 100 Watt  electrical current through the gel. Apparently the DNA is supposed to show up using gel electrophoresis. I think the only error that may be made was pipetting such small amounts of the primers into the tubes. I feel like it was hard to tell if your pipette was actually extracting and releasing the 1 μL of COX1 primers into the tubes but we will see next class.