Lab 7 PCR Protocol and Gel Electrophoresis 2/22/2018

Purpose

The purpose of this lab was to amplify our DNA through the PCR process by denaturing, annealing, and elongating our  samples. After that we started the process of gel electrophoresis by making our mold for next week.

Process

Since the DNA extraction protocol was finished by our TAs (THANK YOU) we started right on the PCR protocol after reviewing the basic steps of what it does. We figured out the amount of DNA, primer, and water we would need in each tube and then mixed all of them. The primers consisted of a forward and reverse primer from the COX1 gene. After all these components were added into the negative control, positive control, and DNA tube we handed them off to be denatured. Group 5s tubes are in B1,2, and 3. The chart of our values is shown in the data section below. After this, We mixed 40 mL of fluid with .6g of agarose gel to make a 1.5% concentration mixture of gel for our electrophoresis mold. We microwaved the mixture, placed it in a water bath, and poured the slightly cooled mixture into the tubes. Our tray has a slight dip in the middle that could possible be a problem when we do the electrophoresis.

Data

note the dip toward the back of the gel plate

Conclusion

This lab was one of my favorites so far. I’m excited to see the results of the PCR process. Hopefully, there won’t be a problem with it because we didn’t get the results we wanted from the Nanodrop. We will be able to tell if it effected it by comparing it to our Paramecium positive control. Next we will unfreeze our DNA and start the electrophoresis phase. Group 5’s tray is stored in the fridge with the others labelled “LSKG5”