Citricular Reaction Lab

Four test tubes were labeled “H2O A”, “H2O B”, “Enz A”, and “Enz B”. In the test tubes labeled “Enz A” and “Enz B” 1 mL of catalase was pipetted into each, and in the test tubes labeled “H2O A” and “H2O B” 6mL of deionized water was pipetted into each. Next the two test tubes labeled A were placed into an ice bucket, and the test tubes labeled B were placed in a 50°C water bath. These were left in their respected locations for 5 minutes each, and then the temperature of the test tubes in the ice bucket were recorded. Next the “Enz A” test tube was mixed with the “H2O A” test tube and 100µL of guaiacol and 150µL of .1% H2O2 were added to this mixture using a micropipette. The test tube was then covered with aluminum foil and inverted to mix the solution and then placed in a curvette and into the spectrometer. The initial absorbance was recorded and the curvette was placed back in the ice bucket. Then the contents of the test tubes labeled B were mixed and 100µL of guaiacol and 150µL of .1% H2O2 were added to this mixture using a micropipette. This was then placed in a curvette and into the spectrometer and the initial absorbance was recorded and the curvette placed back in the water bath. Finally after 5 minutes of time had passed each of these were placed in the spectrometer again and their absorbance levels recorded. This process should be a very quick one to ensure that the

The a-Ketoglutarate undergoes oxidative decarboxylation, loses a CO2 molecule and is then catalyzed by a-ketoglutarate dehydrogenase complex and becomes succinyl-CoA. In the process, a NAD+ molecules becomes an NADH molecule.

As mentioned previously in this paper, the formazan is the compound that is absorbing the light, which means that at a higher absorbance there is more formazan in solution. In order to produce formazan, there must be live, viable cells that can convert MTT; thus, at a higher absorbance, there are more viable cells able to convert MTT. The highest absorbance value is shown to be from the lowest concentration of aspartame, 0mM, which means that this concentration at the highest number of viable cells. The downward trend show in the graph indicates that there are fewer viable cells at each increasing concentration, meaning that as concentration of aspartame increases, cell viability

Objective: To determine if mass has increased or decreased during a chemical reaction. Hypothesis: There will be no change in mass for reaction one and each reactant. From the information of the Law of Conservation of Mass, there will not be any change in mass during and after the chemical reaction. There will be no change in mass for reaction two and each reactant.

In this lab, the study of the kinetics of a chemical reaction will be investigated. One of these reactions involved the oxidation of iodide ions by bromate ions in the presence of an acid.

Table 3 The Averages Absorbance of The Cell Lysate & Cell Pellet In E.coli Induction Experiment

Rat liver cells can be separated and analysed by Subcellular fractionation using homogenisation and then centrifugation to separate the cells into the subcellular fractions. When the fractions are separated succinate dehydrogenase is used to identify mitochondria. Succinate dehydrogenase is an enzyme that catalyses the oxidation of succinate to fumarate in the Krebs cycle and is usually on the mitochondria inner membrane and not found in other organelles in the cell(Rustin, Munnich, and Rötig, 2002). Care needs to be taken when separating the subcellular fractions as organelles can be damaged and can cause lysis giving incorrect results. Regulation of conditions like the temperature and the pH are important as incorrect conditions could cause the enzymes to denature. Also adding an isotonic buffer not water will help to avoid damage to cell organelles due to osmotic an imbalance (Greenawalt and Vasington, 1968). When it comes to measuring the SDH it is measured indirectly because the succinate dehydrogenase and the fumerate involved in the reaction are both colourless so it is impossible to measure any changes using a spectrophotometer. Instead the absorbance of formazan is measured at 490nm. The formazan is produced by reacting the FADH2 from the SDH reaction with INT which a tetrazolium salt. The formazan produced is red in colour and when dissolved into the ethyl acetate can be extracted and then read on the spectrophotometer (Smith and McFeters, 1997). As

Once lab began, the IPTG was added to the time 0 samples cultures, a final concentration of 1mM was added to both cultures, and the timer was started. Every 30 minutes over a time period of 90 minutes 2ml was removed from each culture and the initial flask was returned to the 37° environment to continue growing. The 2ml was equally divided into two different test tubes. The tubes were labeled A and B. There was four tubes at each time point; 2 for lacZ+ and 2 for lacZ-. Tube A was the control tube, which contained .2ml of water. Tube B was the experimental tube, which contained .2ml of water and .013M ONPG. The tubes were then incubated in 37° bath water for 20 minutes. After 20 minutes elapsed the tubes were removed and 2.7ml of 1M Na2CO3 was added to each tube. Next 1ml was transferred from each tube into a cuvette, which was placed in the spectrophotometer at 420nm. A spectrophotometer, also known as a spec, reads the intensity of light (absorbance level of liquids). Tube A was inserted first in order to blank the spec, and tube B was second, so we could record the absorbance of the solution. The absorbance was recorded for each cuvette containing a solution from tube B. The A420 recordings we were needed to determine ONP produced in mmol and the activity. The formula for activity is ONP produced divided

In this experiment we wanted to observe the effects that different types of solutions have on living cells. The three types of solutions that we were looking for was: isotonic solution………

For our cell counting lab, we used three tubes and label them tube #1, tube #2 and tube #3. Once done we had to take 1ml of cell solution to add to tube #1, in tube #2 we added 0.9 ml of buffer and 0.5ml of the buffer to tube #3. I then took 0.1 ml of cell solution from tube #1 and added to tube #2; to mix we inverted our tube four to five times. 0.5ml of cell solution was taken from tube #2 and added to tube #3 once added we inverted the tube to make sure it mixed well. After mixing the tube I took 10ul of cell solution from the tubes and added it to the hemacytometer, to count our cells in the four 16 squares boxes under our microscope, I repeated the last step for all three tubes. Once our cells were counted, the total number of cells was recorded. Tube#1 cell count was very high it showed 603 cells that were counted under the microscope using the hemacytometer. Tube #2 cell count was low but it showed 299 cells. Tube #3 showed hardly any cells under the microscope 22 cells was counted. After getting the total

The result is the high enzyme concentration had the fastest the catalytic

In 1840, there is a book that include observations of mitochondria. In 1894, Richard Altmann called cell organelles to “bioblasts”. In 1904, Friedrich Meves wrote the first record about observation of mitochondria

This ensured that our experimental samples were only showing readings for the light absorbed by the DPIP and not by the mitochondrial suspension as well. Tube 1 consisted of 4.4 ml buffer, 0.3 ml DPIP, 0.3 ml mitochondrial suspension, and 0 ml of succinate. This served as a control because there was no succinate to give off electrons and reduce the DPIP. Tube 2 consisted of 4.3 ml buffer, 0.3 ml DPIP, , 0.3 ml mitochondrial suspension and 0.1 ml succinate. This was the trial with a low concentration of substrate. Tube 3 had 4.2 ml buffer, 0.3 ml DPIP, 0.3 ml mitochondrial suspension and 0.2 ml succinate. This final tube was used to notice the effect of increased substrate on the amount of electrons given off, or the amount of DPIP reduced. We also made sure to add succinate to each tube last, so that the succinate to fumerate reaction would not occur before the DPIP was there to accept the electrons. Immediately after creating each cuvette, we covered it in parafilm to minimize contamination and placed it directly in the spectrophotometer to get an accurate reading. We recorded the absorbance of cuvettes 1, 2, and 3 every 5 minutes for the next thirty minutes and recorded the absorbances in a table. We also made sure to properly mix the contents of the cuvettes thoroughly before getting a reading in order to best distribute the DPIP throughout the solution.

Introduction: - The mitochondria is a double membrane organelle that is found in all eukaryotic organisms. There is some evidence that proves some eukaryotes lack mitochondria, but there is no true evidence about complete lack of mitochondria in the organisms. Mitochondria is known as a power house of the living body cell. They store energy in the cell and release it as needed. Every structure in Mitochondria have their own specific roles which helps in storing energy (See Figure 1 for details). Evidence shows that mitochondria evolved from primitive bacteria. Is it really evolved from primary bacteria or were they the new adaptation in the plant and animal body? In this paper we are going to see how Mitochondrion have been evolved in different species and what methods have been used to prove that evolution. There is still debate between scientists about whether mitochondrion really evolved from bacteria or if it was present in the cells before bacteria were present. Yung et al. [1] says that Mitochondria’s cytochromes c (which is a mitochondrial intermembrane protein that is loosely attached to the inner membrane of mitochondrial membrane) is relatively close to the bacterial medium subunit in sequence of cytochrome. However,

Antioxidative activity can be tested using total phenolic content (TPC), 2, 2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP) tests. In TPC test, Folin–Ciocalteu reagent (mixture of phosphomolybdate and phosphotungstate) is used to determine the antioxidant potential of phenolic and polyphenolic from the sample. DPPH bonds with hydrogen atom from sample while hydrogen peroxide (H2O2) bonds with iron ion (Fe3+) to form iron (III) thiocyanate, Fe(SCN)3 in the FRAP test. The phenolic acids and flavonoids are contributing to antioxidant activities. Radical scavenging activity of petroleum ether extracts of C. nutans was 82.00 ± 0.02 % (Arullappan et al., 2014, pp. 1455-1461). Chloroform leaves extract of C. nutans