By Lauren Buysse, Eli Landas, & Aidan Forberg
Introduction:
The Cancer and Immunity Research Stream here at Mankato State University is focused on how the genes APOBEC3A and APOBEC3B connect to certain proteins and how they lead to the development of cancer. MiRNA are small RNA molecules that can affect how the genes, such as those for APOBEC3A and APOBEC3B, are expressed. So far in the lab, we have followed protocols in order to ensure that we have a solid control to use for the remainder of our research.
Research:
So far in the lab, our group has had very positive results. We have so far taken cells and reduced them down to as high of a concentration of DNA as possible. With that isolated DNA we measured it in order to get the data to figure out how well we did with our DNA isolation. It came out to be approximately 60 microliters per microgram which is well within the desired range (0.2-200 microliters per microgram). With the success of our DNA isolation, our DNA sample was ready to be PCR amplified and put through an Agarose Gel. This “Gel Electrophoresis” gave us the ability to see the size in base pairs of the DNA we had amplified. Our lab for this was successful and gave us the thumbs up that we have followed all of our protocols correctly and gave us the go ahead to start on our actual research next week.
In order for us to move on with our research, we used a database that gave us insight into which MiRNAs had the highest target score and would most likely lead to successful results (them binding to one of the APOBEC genes). The two genes we chose are hsa-MiR-4314 and hsa-MiR-4786-3p. They had target scores of 96 and 90. These target scores are high so we expect that they will bind with the APOBEC gene. Using a couple different online websites and applications we got all the information we needed to order the specific things needed in order to move forward with our research.
Reflection:
Overall, our research stream has been moving at a steady pace. We are able to get into the lab once a week for the time needed to complete each of our experiments. Although we have not conducted any new research yet, we are excited to begin trying out our MiRNA and seeing the results over the coming weeks.
