Who Is Diseased? No one... That we can tell

Our lab took place over three days. The first day consisted of us making our samples with our own cheek cells, after collecting them in flip-top tubes, we then put them into water baths at 56 degrees Celsius for five minutes, vortexed them, then placed them back in the water bath for a remaining five minutes. After finishing the designated ten minutes, we transported the samples to a 100 degree bath. After five minutes we centrifuged and then put them in the refrigerator until the next day.

The second day of the lab we performed PCR amplification. We retrieved our samples from the refrigerator and then we transferred 20 micro liters of the DNA template from the screw cap tube into the bottom of the PCR tube. We were very careful in not transferring any of the matrix beads because they would have killed the DNA polymerase. We added the yellow master mix to each of our samples and mixed well. Then we added our PCR tubes into the thermal cycler, where it underwent 40 cycles of PCR amplification. 


On the third day, we retrieved our tubes and attempted at performing gel electrophoresis. We filled ten lanes of the gel, the first four with "controls" used to compare our results with. The second four lanes had our samples in them. But, unfortunately when we were adding our results one of the samples was being added when the bottom of the gel was punctured and the sample was flooded underneath the gel. Unfortunately, when we got our results back none of our samples showed up except the controls and my own sample (which was the first loaded). So we could not compare our results to our controls to find out who had this diseased gene. However our one sample that did show up, my own, was heterozygous. The way we know this was because after the gel electrophoresis was performed my sample and our heterozygous (+/-) control lined up with my sample. 


Although our results did not all show up, we did learn how to test for certain genes and I was able to see how this process could be important to scientists and other people trying to compare information in different genes.