Lab 1 Report

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School

University of Iowa *

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BIOL:1411:

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Biology

Date

Apr 3, 2024

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pdf

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7

Uploaded by AdmiralEmuMaster1095

Lab Report 1 Arya Sajadian, Ian Johnson, Tom Ohl, Thomas Harold Section 0A09 20 Feb 2024
1 Experiment 1 (Parts I and II) Question and Hypothesis The question posed by this lab was to test for mitochondria in cell fractionates that were separated based on mass via centrifugal force. In order to test how much mitochondria was present, an artificial electron acceptor, DCIP, was used to show a visible change in any mitochondria undergoing the citric acid cycle. The DCIP loses color due to being reduced from SDH-FADH transferring electrons to it, therefore indicating the presence of mitochondria. The hypothesis postulated is that pellet 2 would have the most mitochondria, due to the fact that most of the organelles did congregate there, but did not contain any larger structures like nuclei. Results
2 The goal of the experiment was to see how to observe organelle functions such as the citric acid cycle, understand how the internal workings of the cell are able to be divided up, and be able to see how electron reduction and oxidation occur with SDH’s reduction into SDH-FADH and the subsequent oxidation, as the electrons get transferred and reduce DCIP. The general steps that were taken were to first prepare and divide a cell homogenate, then to prepare several cuvettes to monitor the absorbance. Four cuvettes were used to monitor DCIP absorbance reduction with no homogenate, pellet 2, pellet 3, and supernatant 3, and an additional 4 were used as blanks that contained no DCIP. The data from this experiment showed some initial change in the blank, but little throughout, which can be attributed to a slight error in being unfamiliar with the Spec-200, leading to that discrepancy. Pellent 2 had some notable descent in absorbance, totaling in a 0.08 difference. Pellet 3 had significant descent, totaling in at 0.17 absorbance difference. Supernatant 3 exhibited no change whatsoever throughout the experiment. On the whole, there is not much of a trend to follow due to the small sample, although one could argue a possible bell curve shape, with pellet 3 having significantly more than both other “sides”, being pellet 2 and supernatant 3.
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