Macromolecular Analysis

Figure 1 contains gel electrophoresis for protein samples. The lanes were labeled from 1 to 10 from the right to the left. Lane 1 contained the ladder fragment. Lane 2 contained the filtrate. Lane 3 contained the S1 sample. Lane 4 contained the P1 sample. Lane 5 contained the P1 medium salt sample. Lane 6 contained the P1 high salt sample. Lane 7 contained the S2 sample. Lane 8 contained the P2 sample. Lane 9 contained the P2 medium salt sample. Lane 10 contained the P2 high salt sample.

This technique separates Rubisco samples based on their size. The electrophoresis has a positive and a negative end. Positive charge proteins are loaded from the positive end and migrate towards the negative end. Negative charge proteins are loaded from the negative end and migrate towards the positive end (Sakthivel & Palani, 2016). The sample that contained the highest molecular weight of Rubisco will travel the shortest distance on the gel while the protein with the smallest molecular weight will travel the longest distance (Sakthivel & Palani, 2016). The size proportion of each Rubisco molecule correlates with the distance traveled. Rubisco will be in its purest form after running through SDS-page since each technique will increase the purity of the protein. If the salting out, the ion exchange and the SDS-page protein isolation techniques are performed on protein Rubisco, then it is purified and separated by solubility, charge, and size. The rationale of this experiment is to isolate the purest form of Rubisco so that it can perform carbon fixation at an optimal

We placed the gel into the running chamber, and then we completely covered the gel with TAE. 3 microliters of loading dye was added to each tube; this would help distinguish the enzyme from the gel. As before, we tapped the tube on the table to mix. Then we carefully added each of the four samples into their own wells. A total of 33 microliters of each sample was poured into each well. Afterwards, we attached the positive and negative electrodes to their corresponding terminals on the power supply and gel box. We turned on the power to around 80 volts and waited 45-60 minutes for the loading dye to move down the gel approximately 6-8 cm. Finally, we were able to visualize the DNA in the gel and write down the

Proteins are important elements in cellular membranes and give the membranes many of their characteristics. In red blood cells, the meshwork of proteins in and around the membrane gives it strength and flexibility, allowing a cell to squeeze through small capillaries without bursting. Other proteins play roles in transporting material in and out of the cell (Lab Manual, Cell Biology). Polyacrylamide gel electrophoresis (PAGE), with all of its different modifications is probably the most widely-utilized procedure in contemporary biochemistry and molecular biology (Mordacq and Ellington (1994)). In this experiment, we will attempt to determine the molecular weights of the major proteins in the plasma membranes of bovine red blood cells (RBCs). The predictions made are if our protein has similar weights as proteins

Biomolecules are present in every living organism. Larger biomolecules (macromolecules) consists of proteins, lipids, and carbohydrates. In layman’s terms, proteins are long chains of amino acids that have many functions such as, providing structural support and regulating many body processes. Lipids are the scientific term used for fats. There are many different types of fats, but they all share one common characteristic: They are not soluble in water. Lipids provide protection and insulation to organs and also act as an energy source. Carbohydrates are made up of carbon, hydrogen,

Title: The Hunt for Carbs, Lipids, Proteins and More Statement of the problem: In this lab the main question we have been trying to answer is where did our victim have his last meal? To answer this, we first have to answer questions like: What did his stomach contents contain? And: What does this mean? Answering those questions is tricky, especially without any prior knowledge of macromolecules.

1. Carbohydrates: Are Molecule composed of carbon, hydrogen, and oxygen, and they include sugars and starches.

The controls in this lab are distilled water and baking soda solution because they are tested in every part of the lab. It is important to have these controls for each test because they do not contain any macromolecules so they should test negative for carbohydrates, proteins, and lipids, and providing a base example of a negative reaction. This is useful because it is known they should not contain any of the macromolecules, so if they reacted, it is possible cross contamination occurred. Knowing whether cross contamination occurred allows students to examine their results more carefully.

This experiment overall was very successful as using information obtained from the results of both the single and double digests, a credible restriction map for the unknown plasmid could be constructed. Within this experiment, both single digest and double digests consisting of three restriction endonucleases were used in order to map out the restriction sites of the enzymes making up an unknown plasmid. In order to separate the DNA fragments by their distinct number of base pairs, it was necessary to run an agarose gel electrophoresis. For this particular experiment, a 1% agarose gel was used as this concentration ultimately results in pores that can separate the DNA by size. The process of gel electrophoresis is made possible by the electric current that is used to move the samples of DNA throughout the gel. For this particular experiment, the gel was run at a current of 100 volts. As a result of the phosphate backbone of DNA, DNA is negatively charged. Because DNA is negatively charged, it moves away from the negative electrode and moves down the gel toward the positive electrode as it is attracted that way. What allows for the resistance the fragments face when moving away from the negative electrode is the texture of the gel itself. As a result, the slower fragments of DNA have the ability to move at a faster rate than the larger fragments of DNA. Each sample loaded into the gel contains a loading-dye for two reasons. One reason is that the loading dye contains

SDS-PAGE separates proteins according to their size. Sodium dodecyl sulfate (SDS) dissolves hydrophobic molecules and carries a negative charge. A cell incubated with SDS would have its membranes dissolve and all the proteins becoming soluble and covered with negative charges. As a result, all the proteins only have their primary structures and a large negative charge allowing them to migrate towards the positive pole of an electric field. Polyacylamide gel is used to separate proteins according to their sizes. The gel has pores of different sizes that act as obstacles for the proteins to pass through. Switch off the current, stain the proteins, and the end result would be bands of protein separated according to their molecular weight. One

The support medium for electrophoresis can be formed into a gel within a tube or it can be layered into flat sheets. The tubes are used for easy one dimensional separations, while the sheets have a larger surface area and are better for two- dimensional separations.

When studying macromolecular interactions in vivo, it is essential to use methods that rapidly ‘‘freeze’’ these interactions as well as prevent the re-assortment of protein and RNA components during cell lysis. Cross-linking agents have been exploited for this purpose. In particular, crosslinking agents that are reversible are the most useful because they simplify subsequent characterization of the interacting molecules. Cross-linking to stabilize RNA and protein complexes has proven to be a useful tool for site-specific interactions. Such linkages can generally be achieved by UV or chemical means (Pashev et al., 1991; Matsunaga et al., 2001). The interactions or mere proximity of macromolecules can be studied by the clever use of crosslinking

The purpose of this investigation was to identify the class of immunoglobulins using, sodium dodecyl sulphate (SDS) Polyacrylamide Gel Electrophoresis (PAGE); by making deductions about the structure and molecular organisation of the protein. The experiment was conducted to calculate the unknown molecular mass for reduced and non-reduced immunoglobulin, using SDS-PAGE by measuring the distance migrated. Even though the method is intrinsically inaccurate, it’s enough to deduct the class of immunoglobulin using Mr The experiment displayed that the distance migrated decreased as the molecular mass (Mr) increased. Based on the number of bands visible on the gel, it was concluded that sample A was reduced with 2 bands; heavy chain with Mr of 52.939 kDa and light chain with Mr of 23.470 kDa. Whereas the B sample was non-reduced, with a total molecular mass of 152.415 kDa. This illustrates that both reduced and non-reduced immunoglobulins are of class IgG.

Perform a series of accurate tests on biological molecules to detect the presence of carbohydrates and proteins, as well as the action of an enzyme on specific molecules.

In the laboratory, we are testing the DNA of our two suspects, and the DNA sample from the crime scene. My lab assistants and I will be using an agarose gel to analyze the DNA. We will first dissolve the agarose powder by heating it in a buffer, and then pouring it into a casting tray to harden. Small pores will form as the gel hardens, and they will act to separate the DNA samples. Next, the cooled gel is placed into an electrophoresis chamber. It will be covered by a 1xT TAE buffer. We will load the DNA samples into the compartments and the electrophoresis chamber will be plugged into a power supply. When the power is connected to the chamber, electric currents will move through the agarose gel. The DNA fragments are negatively charged, so when placed into an electric field, they will be attracted to the