Purpose:

• The objective of the lab was to prepare the DNA for amplification. by making a positive and negative control along with a soil sample one. Also we started the process of gel electrophoresis by creating the gel agarose and storing it for the next lab once we continue.

Procedure:

1. Before we began the process, we made sure to wash our work area with 10 % bleach
2. The researchers were given a sheet that had a table of values of the proportions of water, primers, 2X master mix, and DNA we need to pipette into a mini tube.
3. These values were different for each of the groups that we had: positive control, negative control, and our soil sample.
4. Once we included the respective proportions of contents in each of these tubes, we made sure to label the mini tubes with our group number and placed on the rack with the rest of the class.

Preparing the Agarose Gel

1. First we determined how many grams of agarose powder we need for a 1.5% solution of agarose T.A.E. by multiplying 1.5 with 40 mL of T.A.E.
2. We determined the mass of agarose needed was approximate 0.6 grams.
3. The 0.6 grams of agarose powder was then transferred to an Erlenmeyer flask and 40 mL of T.A.E. was then put in the flask also and sealed by wax paper.
4. The flask was then swirled and inserted into a microwave where it was in there for 1 minute and 20 seconds under 70 power.
5. After it was taken out of the microwave, we carefully placed it in a water bath for 5 minutes.
6. While the solution was in the water bath, we assembled the gel rack.
7. Once we took the flask out, ethidium bromide was added and swirled in the agarose gel.
8. We then poured the solution into the gel rack and labeled it with our group number.

Data

 Conclusion:

• This lab allowed be to attain a better understanding of how to make the gel used for gel electrophoresis and how primers can be used in polymerase chain reactions. Also, how specific regions in DNA can help us code for what type or organism we are dealing with.

Further work:

• Next class we will amplifying these cells and then learn how metabarcoding will be carried out.