November 18,2016

The purpose of this lab is to observe the different bands of DNA that we extracted from previous labs.

Procedure:

Making Gel:

1. Use 20 mL of 50xTAE stock solution and 1,000 mL of DI water to make 1xTAE solution
2. Add 40 mL of 1xTAE solution to 20 grams of agarose in Erlenmeyer flask
3. Cover with weighing paper and loose fitting cap
   1. DO NOT COVER TIGHTLY
4. Heat so that the solution becomes clear
5. Let cool for 5 minutes
6. Add 2 mL of ethdium bromide and gently swirl

Setting up electrophoresis box:

1. Set up gel electrophoresis box and make sure the open ends are sealed
2. Pour agarose gel into mold with as little air bubbles as possible
3. Insert comb at one end of the gel
4. Allow gel to set for approximately 30 minutes.
   1. While setting, cover gel with 1xTAE buffer solution so it doesn’t dry out
5. Remove comb and move gel so that the wells are on opposite side of positive electrode
6. Use a micropipette and add 5 mL of the ladder and 10 mL of each PCR product ad loading buffer into the wells
   1. Be careful to not puncture the wells so that the DNA does not fall to the bottom of the gel
7. Place lid on box and turn on the power source to about 100 volts and let gel run until the dye has reached half way through the gel

Conclusion:

There were not any DNA bands to be found in our gel when we took a picture of it using the UV imaging device. This result was disappointing because we had thought that we had done a good job extracting the DNA; however, we were not the only group with this result. If time was not a problem, I would have liked to attempt to extract more DNA from the soil sample using the other extraction technique and tried the gel electrophoresis again.