Aaron Oberstadt

The majority of our group's problems match those of others in the class. We decided to repeat the fish protein lab with various mollusks. This proved to be difficult to exactly replicate in our second gel as the mussel samples had all died. Like Asmah mentioned, we had significant bubble production when transferring Laemelli buffer. Any major issues with bubbles or liquid that would not settle to the bottom of a tube, we found, could be solved with a centrifuge. Finally, on our second gel, a large number of our protein bands were extremely faint, which made reading it very difficult.

These issues, among others, will be addressed in our error analysis. Lacking a second sample of mussel is potentially a major cause of error in our final results, and we could not have prepared for that. We did not maintain consistency in who transferred the protein samples from their respective tubes to the polyacrylamide gel, which could be a source of error. The second gel ran for 5 minutes longer than the first gel, potentially explaining the faintness of its protein bands. Finally, our inability to measure bands that should exist but are too faint to accurately see can also contribute to error in our results.

Research Project Blog #2

My group's goal is to see how well the muscle proteins within bear, caribou, deer, elk, moose, and chicken legs differentiate from one another. Our hypothesis is that, while the proteins of all four species of the deer family will be fairly similar to each other, deer and elk will be most closely related to each other, and caribou and moose will be the closest related to each other as well. However, we found the surprising result of bears having very similar proteins in their muscle tissues as the other mammals! I suppose it's not TOO surprising, since bears are also mammals but the results are actually very identical. (see below) As a matter of fact, all the sampled animals have relatively similar proteins in their muscles, with chicken being the one who holds the most difference.

Lane components:

1. Empty
2. Kaleidoscope standard
3. Elk
4. Moose
5. Bear
6. Chicken leg
7. Caribou
8. Deer
9. Actin & Myosin standard
10. Empty

Figure 1. Result of the first gel electrophoresis.

Figure 2. Result of the second gel electrophoresis.

Due to this, we're faced with greater challenge of distinguishing the differences between each muscle tissue. The bands on the second gel are also smeared in many places, most likely due to too much muscle tissue in the sample or poor pipetting technique, making it even more difficult to read the results. There could also be contaminants on the razors that would interfere with our data, all of which will be discussed in the error analysis section of our project. I'd like to advise other groups to choose a variety of species for more interesting results, and to avoid cutting too much of the muscle tissues when preparing your samples for gel electrophoresis. Less is more!

Research Project Individual Blog Post #2

Individual Blog Post #2
Asmah Tadmori

The challenge that encountered our group is the presence of some smearing in our bands that makes it harder for us to detect which is a true band and which is not. Smearing problem can't help us identify the relationships between our mammals and chicken easily in our phylogenetic tree. Another challenge was while pipetting the buffer using a plastic pipette. We couldn't know if we loaded the appropriate amount of buffer due to the presence of bubbles during each time of pipetting. My question for other people is that, Do you guys experience any bubbles while you were pipetting buffer? And Did you have a smeared bands that made it difficult for you to identify the similarities between the mammals?

For my error analysis, I would assume that contamination has occurred especially while we are using the edged razor blade because we might hold it different ways, and our hands may hold some bacteria. Specifically while cutting the meats, the meat piece start touching most sides of the cup, sometimes the edges which the place where hands touch it. Furthermore, we didn't heat the Actin & Myosin standard at the first gel running, whereas, in the second gel running, we heated it because we didn't know if we had to heat it or not. Consequently, I will discuss the contamination of our skeletal muscle samples, and heating Actin and Ayosin standard sample consequences. Additionally to the movement of our gel that might lead one of the samples in the gel to spill out and cause smearing.