30 January 2023
Emma Rieper & Anna Wilcox
These past two weeks we have been performing PCR reactions and gel electrophoresis. We’ve been becoming more comfortable with lab processes such as pipetting small amounts, making gels, and photographing our results.
We started the week off by learning the basics of how to make and set up PCR reactions. First, we had to calculate the volumes for the correct number of reactions that we would be performing. We then pipetted these volumes into test tubes. DNA was added to one of the tubes to create a positive control, and water was added to the negative control. We then ran these mixtures through a machine called a thermocycler. A thermocycler changes the temperature of the mixture periodically to break down the DNA samples. We then used these mixtures in the gel electrophoresis experiment.
To begin our gel electrophoresis experiment we weighed and measured the agarose gel and the 1x TAE buffer solution. This was then swirled and microwaved to dissolve the agarose gel completely. Making sure the mixture was only slightly cool, we ran it under cold water for a few seconds, swirling the entire time. We then pipetted 10 microliters of the Bullseye dye to our solution and poured it into the gel caster. This gel took approximately 30 minutes to solidify. After it was solidified, we removed the comb that imprints wells into the gel, this is where our samples will go. The gel was then placed into the gel machine. We pipetted liquids into three wells. These liquids were a ladder and our positive and negative controls. We then adjusted the volts on the machine to match our instructions and ran the gel.
After the gel had run for approximately 30 minutes we removed it and brought it to the gel imager. We placed the gel into the imager, followed the protocol, and took photos of our results.