Determining phylogenetic relationships of marine invertebrate and vertebrate using variation in muscle proteins of different genera; Ostreidae , Teuthida, Pectinidae, Nephropidae, Caridea, Brachyura and Oncorhynchus.
Introduction
The fundamental core of all biology emanates from evolution, as the great evolutionary biologist Theodosius Dobzhansky once said "Nothing in biology makes sense except in the light of evolution"(1973). Therefore, studying the phylogenetic relationships of different organisms offers numerous advantages that go well beyond those of just pure research. Modern scientific literature states, that the last common ancestor of vertebrates and invertebrates had muscle. Additionally it is also well documented that most vertebrate …show more content…
We acquired samples of the 5 genera of invertebrates; Ostreidae (Oyster) Pectinidae (Squid), Nephropidae (Lobster), Caridea (Shrimp), and Brachyura (Crab) and acquired 1 genus of vertebrate Oncorhynchus (Salmon), from the local food markets, all samples were froze and stored. For the experiment, 250 µl of Bio-Rad Laemmli buffer was added to a small piece of each sample, the samples were then agitated and allowed to incubate for 5 minutes at room temperature. The variation in different sample protein structure was measured solely on the bases of sizes (molecular weight), and no other physical property such as charge. Therefore all protein samples were first denatured by placing them in 95°C for 5 minutes, so that they no longer possessed any secondary, tertiary or quaternary …show more content…
The pre-cast gel was placed in a Pierce Precise Protein Gel Cassette. The gel was an 8x8 mini, 1mm thick and had 10 wells for loading samples. The inner chamber of the cassette was completely filled to the brim with 150 ml of Tris-Glycine gel running buffer, the mini tank outside was filled with the same buffer, to approximately 1/3 of the container.
The protein samples were then placed in each of the wells, in an order than can be seen in Figure 1. The first well was filled with a protein standard to identify protein separation during gel electrophoresis and to estimating the molecular weight of sample proteins. The gel was run for an hour in at a voltage of 108V, after which the gel was stained using Coomassie Blue dye. Figure 3 represents an image taken after the gel was stained and rinsed in distilled water.
Introduction
The fundamental core of all biology emanates from evolution, as the great evolutionary biologist Theodosius Dobzhansky once said "Nothing in biology makes sense except in the light of evolution"(1973). Therefore, studying the phylogenetic relationships of different organisms offers numerous advantages that go well beyond those of just pure research. Modern scientific literature states, that the last common ancestor of vertebrates and invertebrates had muscle. Additionally it is also well documented that most vertebrate …show more content…
We acquired samples of the 5 genera of invertebrates; Ostreidae (Oyster) Pectinidae (Squid), Nephropidae (Lobster), Caridea (Shrimp), and Brachyura (Crab) and acquired 1 genus of vertebrate Oncorhynchus (Salmon), from the local food markets, all samples were froze and stored. For the experiment, 250 µl of Bio-Rad Laemmli buffer was added to a small piece of each sample, the samples were then agitated and allowed to incubate for 5 minutes at room temperature. The variation in different sample protein structure was measured solely on the bases of sizes (molecular weight), and no other physical property such as charge. Therefore all protein samples were first denatured by placing them in 95°C for 5 minutes, so that they no longer possessed any secondary, tertiary or quaternary …show more content…
The pre-cast gel was placed in a Pierce Precise Protein Gel Cassette. The gel was an 8x8 mini, 1mm thick and had 10 wells for loading samples. The inner chamber of the cassette was completely filled to the brim with 150 ml of Tris-Glycine gel running buffer, the mini tank outside was filled with the same buffer, to approximately 1/3 of the container.
The protein samples were then placed in each of the wells, in an order than can be seen in Figure 1. The first well was filled with a protein standard to identify protein separation during gel electrophoresis and to estimating the molecular weight of sample proteins. The gel was run for an hour in at a voltage of 108V, after which the gel was stained using Coomassie Blue dye. Figure 3 represents an image taken after the gel was stained and rinsed in distilled water.