Rubisco Lab Report

Colorimetric assay is a process of determining a concentration of a solution based on absorbance of light. The purpose of this lab is to determine if the Bradford assay is an accurate way to determine an unknown concentration of two samples of protein. The Bradford assay is done by measuring wavelength of light passing through a cuvette filled with Bradford dye and concentrations of PBS and proteins. After the cuvettes are mixed they are placed into a spectrophotometer to measure wavelength. The wavelength given will be used to plot a standard curve based on concentration (x-axis) and wavelength (y-axis). The standard curve is then used to measure an educated guess on the concentrations of unknown protein concentrations. We hypothesized that if we use the Bradford assay and colorimetric spectrophotometry we can determine an accurate concentration of two unknown concentrations of proteins. The results of this lab failed to reject our hypothesis based on accurate measurements of protein concentrations. The standard curves are drawn with a linear increasing slope. The Bradford assay is an accurate way to demine the concentration of an unknown concentration.

The mole is a convenient unit for analyzing chemical reactions. Avogadro’s number is equal to the mole. The mass of a mole of any compound or element is the mass in grams that corresponds to the molecular formula, also known as the atomic mass. In this experiment, you will observe the reaction of iron nails with a solution of copper (II) chloride and determine the number of moles involved in the reaction. You will determine the number of moles of copper produced in the reaction of iron and copper (II) chloride, determine the number of moles of iron used up in the reaction of iron and copper (II) chloride, determine the ratio of moles of iron to moles of copper, and determine the number of atoms and formula units involved in

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.

The proteins are also added to a Laemmli sample buffer in order to give each protein a negative charge so it is able to get pulled through the polyacrylamide gel. The next step is to put the gel into the electrophoresis module and to run it. It is run until the proteins have almost reached the bottom of the gel. A blue tracking dye is added to the Laemmli sample buffer in order to track the distance in which the proteins travel through the gel. If it is run for too long, the proteins will run off the bottom of the gel and it will mess up your results. Once the protein reach the bottom of the gel, the gel is stained in order to be able to see the individual bands of the different proteins. When the gel is stained, the protein distances will be able to be measured and compared. For a detailed procedure, refer to the Comparative Proteomics Kit I: Protein Profiler Module Lab Manual.

2. When 2.00 g of NaOH were dissolved in 49.0 g water in a calorimeter at 24.0 ˚C, the temperature of the

Our first step was that we boiled the samples, A&M Std , & Kaleidescope Std. for 5min. in water bath, then we loaded the samples into the wells following the guide given which was as follows: 1- 10 ml of laemmli buffer, 2- 10 ml of molecular weight, 3- 10 ml of salmon, 4- 10 ml of tilapia, 5- 10 ml of catfish, 6- 10 ml of shrimp, 7- 10 ml of actin and myosin, and 8- 10 ml of the laemmli buffer. To load each sample, we used a P-200 (yellow) micropipette tip to withdraw 10 ml of each protein sample from its tube and gently transferred it into the designated well. After loading all samples, on both of the gels we then placed the lid on the tank, and insert the leads into the power supply, matching red to red and black to black. We ran the gel for 45 minutes at a constant voltage of 115V. When the gels were finished running, we discarded the buffer from the inner chamber, we released the cams, and removed the gel cassettes from the assembly. We laid each gel cassette flat on the bench with the short plate facing

October 17, 18, and 19, samples were collected from multiple sites along the BSR. The class was split into groups, and samples were collected from seven separate locations along the river and WWTP. There was also a sample collected by the S which is located between sites four and five. For each of these sites, there were ten groups from other labs that also collected a sample from the BSR. At site two of the river, the location included multiple sources of possible contamination. A drainage site was located 200 yards upstream, along with a small PVC drainage pipe next to the collection site. Not only was there drainage running into the river, the site was under a bridge, and contained other trash scattered throughout the area. The

8) Results. What tools did they use to show the localization of the different proteins? What did they find on the localization and function of the different proteins?

Oxygenation in plants wastes energy that could otherwise be beneficial to the plant. A Rubisco engineered to have a higher tolerance to heat would produce a plant with a greater yield potential due to its higher photosynthetic efficiency and would probably have a higher catalytic rate (Parry et al., 2010). Some species of Rubisco has evolved for higher carbon environments like the non-green algae Griffithsia monilis. If C3 plants can express the same Rubisco as this algea carbon canopy gain can be up to 27% leading to higher plant biomass and yields (Long et al., 2006). A Rubisco enzyme with a greater ability specify to CO2 over O2 would lead to a decreased amount of oxygenation of RuBP and a lead to a greater overall efficiency of the plants photosynthesis. Rubisco uses up a total of about 50% of a plant leafs soluble proteins and 25% of leaf nitrogen so increasing the efficiency would lead to more energy being used elsewhere (Lin et al., 2014). Work has been done to increase the carboxylase reactions relative to the oxygenase reaction in Rubisco over the last 20 years by using

The volume was recorded for this solution. The solution was transferred to a beaker and placed into a stirring ice bath, and 1.75 mL of cold methanol was slowly added to the solution for every 1.0 mL of the new volume of SV. After adding methanol, the solution was centrifuged at 10,000xg for 10 minutes. The pellet (PVI) was kept, while discarding SVI after putting aside 1.0 mL for later analysis. Water Extraction of the enzyme from PVI

This procedure involves exposing the enzyme to two concentrations of the salt (35% and 57%) in two different steps. This is called salt fractionation and effectively separates large groups of proteins, lipids, carbohydrates, and DNA. The process works by causing the highly hydrophobic molecules to become insoluble at 35% and the partially hydrophobic (acid phosphatase) to precipitate at 57%, leaving the most hydrophilic solutes in solution.1 The salt concentration and type may be varied based on the isolation compound of interest, for the overlying principle of this technique, selective dehydration, remains the

Thin layer chromatography (TLC) separates compounds, through migration, from a mixed solution with the assistance of a solvent and an absorbent strip of cellulose. The purpose of this lab is to allow students to determine an unknown amino acid by comparing results to six known amino acids (slowest to fastest: lysine, glycine, alanine, tyrosine, phenylalanine, and leucine) and properly separate a combined solution of amino acids using the TLC method. But to fully understand the lab, two terms must be understood: amino acid and protein. Amino acids are building blocks of proteins linked by peptide bonds that contain both carboxyl and amino functional groups.

Part one of this lab consisted of the extraction of onion DNA utilizing a lysis solution. In part two a chain of plastic links was used to mimic the slicing and separating a strand of DNA as restriction enzymes and gel electrophoresis would. In part three, multiple samples of real DNA were compared to each other

For this experiment we also made use of agarose gel electrophoresis, which takes a lot of time. Electrophoresis may be the main technique for molecular separation in today's cell biology laboratory. In spite of the many physical arrangments for the apparatus, and regardless of the medium through which molecules are allowed to migrate, all electrophoretic separations depend upon the charge distribution of the molecules being separated.

His tags are commonly used to purify proteins through immobilized metal-affinity chromatography (IMAC) (Lilius et al., 1991). This rapid and efficient method separates the recombinant protein from unwanted products such as RNA. The His tag DNA sequence is inserted into the