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Introduction: Eukaryotic cells are complex and contain various membrane bound organelles including the mitochondria and the nucleus. A nucleus is a dense membrane bound organelle that contains the genetic materials and the mitochondria are organelles that are responsible for respiration and energy production activity. The Key components and identifiers for DNA are the chromosomes located inside the nucleus. The succinate dehydrogenase, located on the inside of the mitochondrial membrane, is a positive indicator for the presence of mitochondria. Each of the organelles has a specific density. DNA carrying nucleus is the densest organelle followed by mitochondria. Separation of substances with differing density can be accomplished by using centrifugation. Centrifugation uses the centripetal forces to separate two heterogeneous mixtures. For more dense substances lower speeds can separate the heterogeneous mixtures but to remove less dense substances from the solution, the solution needs to be centrifuged faster for a longer period of time. DCIP- The concentration of the solution can be obtained by using spectrophotometer. The higher the OD number, the more concentrated the solution is. It works by measuring the amount of light of a specific wavelength that passes through a medium. Using these properties and techniques separate solutions of mitochondria and nucleus were created and observed Discussion: Under the microscope, purple staining was observed for the F 1 Solution. This Azure C purple staining is indicative of DNA presence meaning that pieces of nucleus were present. The OD values for F 1 at zero minutes were 0.519 and 1.578 at 3 minutes mark. This high OD shows that mitochondrial succinate dehydrogenase.

Intro and Discussion

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cell fractionation

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Page 1: Intro and Discussion

Introduction:Eukaryotic cells are complex and contain various membrane bound organelles including the mitochondria and the nucleus. A nucleus is a dense membrane bound organelle that contains the genetic materials and the mitochondria are organelles that are responsible for respiration and energy production activity. The Key components and identifiers for DNA are the chromosomes located inside the nucleus. The succinate dehydrogenase, located on the inside of the mitochondrial membrane, is a positive indicator for the presence of mitochondria.

Each of the organelles has a specific density. DNA carrying nucleus is the densest organelle followed by mitochondria. Separation of substances with differing density can be accomplished by using centrifugation. Centrifugation uses the centripetal forces to separate two heterogeneous mixtures. For more dense substances lower speeds can separate the heterogeneous mixtures but to remove less dense substances from the solution, the solution needs to be centrifuged faster for a longer period of time. DCIP-The concentration of the solution can be obtained by using spectrophotometer. The higher the OD number, the more concentrated the solution is. It works by measuring the amount of light of a specific wavelength that passes through a medium. Using these properties and techniques separate solutions of mitochondria and nucleus were created and observed

Discussion:Under the microscope, purple staining was observed for the F1 Solution. This Azure C purple staining is indicative of DNA presence meaning that pieces of nucleus were present. The OD values for F1 at zero minutes were 0.519 and 1.578 at 3 minutes mark. This high OD shows that mitochondrial succinate dehydrogenase.

S1 with the Azure C dye had a purple staining under the microscope. This means that the supernatant 1 had traces of nucleus matter. The OD at 0 mins was 0.839 and at 3 mins 0.994. The concentration was less than F1, meaning that the concentration of succinate dehydrogenase in this sample was low. This result suggests that mitochondrial traces were present.

P1 had very few scattered pieces of purple staining in the solution. This staining was observed under the microscope and suggests that there was a very low concentration of DNA i.e. nucleus present. The OD at 0 mins was 1.767 and 1.685 at 3 mins. This observation suggests that there was a very high concentration of succinate dehydrogenase. Most of the P1 solution showed evidence of high mitochondrial concentrations.

S2 had a 0.776 concentration at 0 mins and 0.585 OD at 3 mins. These values suggest very low traces of mitochondrial substances. The microscope created a green picture with what appears to be a large stained area. This might suggest that nucleus material was present.

Page 2: Intro and Discussion

P2 had an OD reading of 0.533 at 0 mins and 0.544 at 3 mins. This observation suggests that there was a very low if any concentration of succinate dehydrogenase i.e. mitochondrial material.

Different concentrations of P2 were placed in a series of solutions (P2A, P2B, P2C, & S2). For P2A the OD reading at 0 mins was 0.955 and at 0.996 at 3 mins. It has a velocity of 1.267 * 10-5. This velocity refers to the rate at which DCIP is reduced. P2B had the starting OD of 0.647 and 0.730 ending OD. The velocity was calculated at 7.90*10-6. P2C had the starting concentration at 0.649 at 0 mins and 0.676 at 3 mins, with a velocity of 2.57*10-6. S2 had the OD reading of 0.749 at the start and the ending OD reading was 0.948 with a velocity of 1.89*10-5. The OD reading for all the values above unexpectedly increased from 0 mins to 3 mins. This error could be because of calibration error of the spectrophotometer. Another potential reason could be very small and limited amount of time provided for reduction of DCIP. It is hard to decipher which concentration of P2 provided the best results for reduction of DCIP based on the data, as all values increased after 0 mark. P1S1