Rahul Palnitkar: Week 8
This past week was my final week at the Seale Lab. It started quite slowly, but the last two days were perhaps the busiest days I had during my time at the lab. So what happened?
Monday was quite a slow day. I arrived at the lab at 8:30 AM as usual, and began working on my poster. I continued working until 10:30, when I realized my supervisor still wasn't at the lab. In fact, almost no one was at the lab except me and two undergraduates. I was a bit confused, but I continued working on my poster. Finally, at 1:00, I left the lab. I later got an email from my supervisor saying that I didn't need to be there that day.
On Tuesday, I had a good amount of work to do. I transfected the virus-infected HCT116 cells with p53 and empty pcDNA plasmids. Besides that, I genotyped 20 more mice, and worked on my poster.
Wednesday was much like Monday, in that it was a slow day. I worked on my poster once again, and genotyped another 15 or so mice. Additionally, I spent about an hour preparing for the next day, when our real work would begin.
Thursday was packed. I don't think I even sat down at my desk once that day! We were doing another co-immunoprecipitation experiment, and we had to rush so that we could finish by Friday, my last day. I must admit, it was quite stressful, but we did get through it. The part of the experiment I dreaded the most was loading the protein gel, as I am rather clumsy. However, to my (and my supervisor's) great surprise, I loaded the wells of the gel without breaking the gel or introducing bubbles to the wells. We let the gel run, and then we started to transfer the proteins to the membrane. Finally, the membrane containing the proteins was left in a mixture of primary antibodies overnight.
On Friday, my last day, we immediately resumed our co-immunopreciptation experiment. We first made some fresh TBST buffer, as we had run out, and then we washed the membrane and stuck it in a mixture of secondary antibodies. Finally, we looked at our western blot under the Li-Cor machine. Unfortunately, we didn't get the result we were hoping for, although that could be due to the fact we infected the HCT116 cells with prdm16 and transfected them with p53, resulting in a much higher amount of p53 and a much lower amount of prdm16.
The bright green lines above are p53, and the bright red lines (except for the far right) are two forms of Prdm16. Before, we showed that Prdm16 interacted with p53. Here, on the other hand, we did not pull down any p53 with Prdm16, which contradicted our earlier results.
After finishing the CoIP, it was time to say goodbye. Two undergrads were also finishing on that day, so we all gathered together and talked and took pictures. Finally, at 3:30, I left the Seale Lab. It was a wonderful experience, and I hope I can go back there next summer.
Monday was quite a slow day. I arrived at the lab at 8:30 AM as usual, and began working on my poster. I continued working until 10:30, when I realized my supervisor still wasn't at the lab. In fact, almost no one was at the lab except me and two undergraduates. I was a bit confused, but I continued working on my poster. Finally, at 1:00, I left the lab. I later got an email from my supervisor saying that I didn't need to be there that day.
On Tuesday, I had a good amount of work to do. I transfected the virus-infected HCT116 cells with p53 and empty pcDNA plasmids. Besides that, I genotyped 20 more mice, and worked on my poster.
Wednesday was much like Monday, in that it was a slow day. I worked on my poster once again, and genotyped another 15 or so mice. Additionally, I spent about an hour preparing for the next day, when our real work would begin.
Thursday was packed. I don't think I even sat down at my desk once that day! We were doing another co-immunoprecipitation experiment, and we had to rush so that we could finish by Friday, my last day. I must admit, it was quite stressful, but we did get through it. The part of the experiment I dreaded the most was loading the protein gel, as I am rather clumsy. However, to my (and my supervisor's) great surprise, I loaded the wells of the gel without breaking the gel or introducing bubbles to the wells. We let the gel run, and then we started to transfer the proteins to the membrane. Finally, the membrane containing the proteins was left in a mixture of primary antibodies overnight.
On Friday, my last day, we immediately resumed our co-immunopreciptation experiment. We first made some fresh TBST buffer, as we had run out, and then we washed the membrane and stuck it in a mixture of secondary antibodies. Finally, we looked at our western blot under the Li-Cor machine. Unfortunately, we didn't get the result we were hoping for, although that could be due to the fact we infected the HCT116 cells with prdm16 and transfected them with p53, resulting in a much higher amount of p53 and a much lower amount of prdm16.
After finishing the CoIP, it was time to say goodbye. Two undergrads were also finishing on that day, so we all gathered together and talked and took pictures. Finally, at 3:30, I left the Seale Lab. It was a wonderful experience, and I hope I can go back there next summer.
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