Bio 1 Lab Report

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University of Iowa *

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1411

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Biology

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Apr 3, 2024

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docx

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BIOL:1411 LAB REPORT 1 Bethel Tewabe Team Members: Gavin Smith, Colten Kuhn, Ashley Jokovich Lab section: 0A29 10/03/23
Experiment 1 Question & Hypothesis The overarching question for the first experiment conducted was: How much mitochondrial activity is present within a specific cell fraction? To do this, we performed cell fractionation of our chosen plant cell and centrifuged them to determine the amount of specific organelles present, but more specifically the mitochondrial activity present with the use of biochemical markers. We hypothesized that our final pellet would have the most mitochondrial activity, most prominent color change, because mitochondria is one of the smallest organelles and with each centrifuge the organelles being filtered out were getting smaller. Results Once the centrifuging was done and we obtained all of our cell fractions, we had a series of cuvettes that received a specific amount of: cell fraction, for measurement purposes, sodium azide, to prevent the transfer of electrons to the electron transport chain, DCIP, which is now acting as the electron acceptor and undergoes a color change when reduced, and succinate, which is the biochemical marker for mitochondrial presence. Once all of these amounts were added to the cuvettes, they were put into the spectrometer to obtain an absorbance reading. Figure 1 Depicted in the graph is the difference in absorbance values at each timed interval.
Figure 2 Depicted in this graph is the overall change in percent value of each cell fraction. Shown in both figure 1 and figure 2, there is no change whatsoever within the E1 cuvette and this is because E1 was the control. Noticeable from figure 1, our E2 cuvette had the largest absorbance reading in comparison to all of the other cuvettes. Only cuvettes E2 and E3 had a significant decrease as opposed to E1 which had none and E4 which had the smallest and, overall, insignificant decrease. E3 had the largest decrease in both graphs, but more prominently seen in figure 2. Discussion Our hypothesis, that P3 would have the highest mitochondrial activity, was supported by our data. This is due to the fact it had the highest decrease in absorbance indicating that the SDH accounted for the presence of many mitochondria in comparison to the other cuvettes. An additional experimental condition could be to repeat this experiment with animal cells and compare data or to implement changes in pH, temperature, substrate/enzyme concentration, competitive/noncompetitive inhibitors which is what we performed in our second experiment (Grewe, 2023).
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