Hello, welcome back to my blog! If you haven’t been following along, my name is Siena Parr and I have just finished my fourth week interning through Scripps Research at the Dorris Neuroscience Center. Last week was a short week due to the holiday but my fellow intern/ roommate Katie and I were back to working five days this week.
Sea Lions!
Monday was a pretty quiet day, we started out the morning with a lab meeting. After the meeting, I made an agarose gel (to use instead of an E-gel) and ran some samples that were leftover from last week. I saved most of the gel for use in the future. I spent the rest of my day neuron-tracing. Our lab had another intern from UCSD start this week and I helped her learn how the tracing software works. After work on Monday Katie and I went to La Jolla Cove and watched the sea lions there before getting cream puffs and going home for the day.
On Tuesday this week, I started off my day with a presentation about microRNAs. There is an auditorium in the downstairs of the neuroscience center and occasionally they have presenters or speakers come and talk about their work. After that, I went over the neurons that I labeled on Monday and then ran a PCR (this was the first one I did by myself, yay!) This specific PCR was run to better determine the best annealing temperature for a few different primers. The past PCRs hadn’t shown strong bands when they were run on a gel and it’s possible that they were being run at the wrong temperatures.. This PCR took around 2 1/2 hours in the thermocycler so I traced more neurons before heading home for the day. My tracing skills and I were deeply humbled by the neuron I traced on Tuesday. It was by far the largest and most structurally complex one I had ever done, I think it took me around 2 hours to complete and I had to go back in the next day and touch up all of the spots I missed.
Primers for a forward primer mix.
On Wednesday I ran an E-gel with the samples I had made during my PCR on Tuesday, along with two of my mentor’s samples. The samples that I made didn’t have strong bands but my mentor’s did and we were able to extract the DNA from them. We cracked the plastic E-gel casing apart and cut out the bands. After that, we dissolved the gel coating in a buffer by heating it until all of the visible solid pieces were dissolved. The liquid was then pipetted into a spin column and centrifuged. We washed the DNA with wash buffer and centrifuged it three more times to make sure the DNA was separated from the agarose. This process was similar to the midipreps that I did the second week of my internship, just on a smaller scale. I also traced more neurons on Wednesday.
Our imaged gel, the ladder is on the left and our samples are on the right.
On Thursday, I ran a gel in the morning, there were no visible bands on it so we couldn’t do the gel extraction that we did yesterday. I also ran another PCR in the afternoon, we almost didn’t get to run this one on a gel since we were out of DNA ladder but we found some! It was also the Jin Lab’s birthday on Thursday and we got bubble tea! Our host family needed to go to Costco on Thursday afternoon so Katie and I tagged along.
Friday was a quiet day, almost everyone was done for the week. I watched my mentor run a qPCR and I mapped another neuron before sitting through an hour and a half of traffic to get home.
Giant Tortoise!
Over the weekend I went to the San Diego Zoo with Katie and some of the other San Diego pinterns. It was a huge zoo and one of my favorites were the giant tortises they had. I’m going to go run on the beach in the morning tomorrow which is exciting and hopefully have a relaxing day!
Thank you for reading, stay tuned for the last two weeks of my internship, it has gone by really quickly. Scripps already sent me my exit survey and I feel like I just got here!
-Siena
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