Part 1: Introduction
Proteonomics, the study of the structures and functions of proteins, is perceived as the "next step" in research, the last step being genomics. The proteome, unlike the genome, differs from cell to cell and constantly changes, so it is fairly difficult to study. Some research suggests that a lot of noncoding DNA in the genome functions to regulate protein production, expression levels, and modifications to proteins after translation, and that these variations are what differentiate humans from other species with similar genomes. We have also learned that one gene can code for many proteins and mechanisms that finely regulate things like cellular locations and expression levels of different proteins. In this lab, we will run several proteins on a gel and determine if they are of similar species.
Part 2: Experiment
First, we willl label one fliptop micro tube and also one screwcap micro tube for each of five fish samples. Then we'll add 250 microliters of Laemmli sample buffer to each fliptop tube. Then we'll cut a small piece of fish muscle from each sample and put it in the appropriate fliptop tube, then flick the tubes to mix it up. Then we'll incubate the tubes for 5 minutes at room temperature. Next we'll pour only the buffer from each fliptop tube into the corresponding screwcap tube. We will heat the screwcap tube samples for 5 minutes at 95 C.
On the next day, we'll set up a Mini-Protean 3 gel box and add electrophoresis buffer to the chamber. We'll prepare the gel as indicated in the lab guide. Then we'll put it in the mini tank, and fill it with electrophoresis buffer. Meanwhile we will have heated the fish samples and actin and myosin standards to 95 C for 2-5 minutes. Then we'll load the gels as indicated in the lab guide, and electrophorese for 30 minutes at 200 volts. Then we'll remove the gels, rinse them off, and stain the gels for at least 1 hour. Then we'll destain the gels in water overnight.
On the third day, we'll dry the gels using GelAir cellophane.
