Rahul Palnitkar, Week 7: Calm before the Storm
This week was a rather short one, as I had Friday off, due to Rachel, my supervisor, not being at the lab. Overall, it was a slower-paced week, which was a welcome reprieve from the extremely packed previous two weeks. I spent a good amount of time working on my poster, but I also did some lab work as well.
The first task I had was to genotype even more mice to see if any one of those mice were triple knockout for Prdm16, p53, and Casp8. At first, I thought I would be let down once again, as in the 60 mice I had previously genotyped, I had not found a triple knockout. However, I was pleasantly surprised to see that not one, not two, but three of the mice I genotyped were triple-knockout mice. This was a great success, as these are the first triple knockouts we've seen in the seven weeks I've been here. However, all of these mice were male, meaning that we couldn't just breed these triple knockouts together to produce more triple knockouts. Nevertheless, it still was a good result.
The week also contained some bad news as well, however. One of my CaCo2 cell plates was found to be contaminated, so I had to spend some time re-plating the CaCo2 cells.
Other than that, I loaded my second protein gel and did my second western blot. This time, we were trying to detect levels of Prdm16 in the colon, the jejunum, and the duodenum, which are all parts of the small intestine. Although the loading of the protein gel didn't go that well, as I am not too experienced at loading polyacrylamide gels, the western blot actually turned out pretty great! Instead of using a developer to visualize the blot, we used a system called Li-Cor, which actually looked better than using film. The blot showed us that Prdm16 was highly expressed in the Duodenum, somewhat expressed in the jejunum, and not expressed in the colon.
Other than that, we did't do much else except prepare for the next week, when we are going to do a second co-immunoprecipitation experiment.
The first task I had was to genotype even more mice to see if any one of those mice were triple knockout for Prdm16, p53, and Casp8. At first, I thought I would be let down once again, as in the 60 mice I had previously genotyped, I had not found a triple knockout. However, I was pleasantly surprised to see that not one, not two, but three of the mice I genotyped were triple-knockout mice. This was a great success, as these are the first triple knockouts we've seen in the seven weeks I've been here. However, all of these mice were male, meaning that we couldn't just breed these triple knockouts together to produce more triple knockouts. Nevertheless, it still was a good result.
The week also contained some bad news as well, however. One of my CaCo2 cell plates was found to be contaminated, so I had to spend some time re-plating the CaCo2 cells.
Other than that, I loaded my second protein gel and did my second western blot. This time, we were trying to detect levels of Prdm16 in the colon, the jejunum, and the duodenum, which are all parts of the small intestine. Although the loading of the protein gel didn't go that well, as I am not too experienced at loading polyacrylamide gels, the western blot actually turned out pretty great! Instead of using a developer to visualize the blot, we used a system called Li-Cor, which actually looked better than using film. The blot showed us that Prdm16 was highly expressed in the Duodenum, somewhat expressed in the jejunum, and not expressed in the colon.
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