Bio Paper

2503 Words Nov 23rd, 2012 11 Pages
Gluskin 6
This experiment attempts to answer the question of whether an increase in a succinate concentration (a component of the Krebs cycle) will lead to an increased rate of cellular respiration within a cell. We measured the amount of electrons given off by the succinate to fumerate redox reaction by using DPIP. DPIP is an electron acceptor that takes the place of FAD by accepting the electrons and turning from its oxidized blue state to its reduced clear state. We had three tubes with varying concentrations of succinate and measured the transmittance of each over a half hour period to determine whether more succinate led to more DPIP being reduced. The results showed that the tube with no succinate (no
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There will also be a buffer added to the solution to ensure that the pH remains relatively constant, preventing the possibility that any enzymes would be denatured. The main question of this experiment is whether or not a high concentration of succinate will increase the rate of the Krebs cycle. My hypothesis is that an increase in the substrate will result in an increase in the rate of reaction. If we increase the substrate concentration, then ore DPIP will be reduced, turning the solution clear and increasing the transmittance.
Materials and Methods: The first thing that we did is flip switch C to turn on the Bausch & Lomb Spectronic 20 five minutes before we started, in order to let it warm up and calibrate properly. We then set the wavelength to 600nm, the wavelength at which DPIP absorbs light. We then used the control knob to set the spectrophotometer by turning the transmittance level to 0%, with the cover closed and the chamber empty. After labeling four different cuvettes B, 1, 2, 3 corresponding to each trial, we prepared the blank (B) solution. Using a micropipette, we measured 4.6 ml buffer, 0.3 ml mitochondrial suspension, and 0.1 ml succinate into the B cuvette that contained no DPIP. Before placing the cuvette in the spectrophotometer, we wiped down the sides and aligned the mark on the cuvette with the line on the holder to ensure that the reading came from

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