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respiration lab. The method is where you show the steps of how to do the lab correctly. The results is how you show the reader of the data that you received, and to compare that with anyone else who wants to do the same lab. The discussion is where you discuss with the reader how your results went, and if any problems occurred, how and why they occurred the way that they did. The conclusion is where you conclude the lab, and allow any other specific types of evidence to further help with the conclusion of the lab. Hypothesis At a warmer temperature, more oxygen will be absorbed by the peas than at a cooler temperature, and germinating peas will absorb more oxygen then nongerminating peas both room temperature and 10 degree Celsius conditions. Introduction In this lab we will measure oxygen consumption during respiration as the change in gas volume in respirometers containing germinating or nongerminating peas. Respiration refers to two different but related processes. Respiration is the active acquisition of O2 by an organism and cellular respiration is the release of energy from organic compounds by metabolic chemical oxidation within each cell. Note the equation below for the complete oxidation of glucose. Oxygen is required for this energy releasing process to occur. C6 H12 O6 + 6 O2 -> 6 CO2 + 6 H20 + 686 kilocalories of energy/mole of glucose oxidized. The chemical oxidation of glucose has important implications in relation to the measurement of respiration. You will measure the: 1. Consumption of O2 during the oxidation of glucose 2. Production of CO2 during aerobic respiration 3. Consumption of O2 and the release of CO2 during cellular respiration In this experiment, the relative volume of O2 consumed by germinating and nongerminating peas at two different temperatures
will be measured. Also the CO2 produced during cellular respiration will be removed in the presence of potassium hydroxide and will form solid potassium carbonate by the following reaction: CO2 + 2 KOH -> K2CO3 + H2O. Therefore the change in the volume of gas in the respirometer will be directly related to the amount of oxygen consumed. In the experimental apparatus, if water temperature and volume remain constant, the water will move toward the region of lower pressure due to the consumption of O2. The vial with glass beads only will permit detection of any changes in volume due to atmospheric pressure changes or temperature changes. The amount of O2 consumed will be measured over a time course. The experiment required six respirometers to be set up as follows.
Method First, both a room temperature bath and a 10 degree C bath should be set up immediately to allow for time to adjust the temperature of each. Add ice to attain the 10 degree C bath. Which the baths are equilibrating, obtain a 100-mL graduated cylinder and fill it with 50 mL of H2O. Drop in 25 germinating peas and determine the amount of water that was displaced. Record the volume of 25 germinating peas. Remove these peas and place them on a paper towel. They will be used in respirometer 1. Second, refill the graduated cylinder with 50 mL of H2O. Drop 25 dried peas into the graduated cylinder and then add enough glass beads to attain a volume equivalent to that of the expanded germinating peas. Remove these peas and beads and place them on a paper towel. They will be used in respirometer 2. Third, refill the graduated cylinder with 50 mL of H2O. Determine how many glass beads alone would be required to attain a column equivalent to that of the expanded germinating peas. Remove
these beads and place them on a paper towel. They will be used in respirometer 3. Fourth, repeat the procedures above to prepare a second set of germinating peas, dry peas + beads, and beads for use in respirometers 4, 5, and 6, respectively. Fifth, to assemble 6 respirometers, obtain 6 vials, each with an attached stopper and pipette. Place a small wad of absorbent cotton in the bottom of each vial and, using a dropper, saturate the cotton with 15% KOH. Make sure that the respirometer vials are dry on the inside. DO NOT get KOH on the sides of the respirometer. Place a small wad of dry cotton on top of the KOH-soaked absorbent cotton. It is important that the amounts of cotton and KOH be the same for each respirometer. Sixth, place the first set of germinating peas, dry peas + beads, and beads in vials 1, 2, and 3, respectively. Place the second set of germinating peas, dry peas + beads, and beads in vials 4, 5, and 6, respectively. Insert the stopper fitted with the calibrated pipette. Place a weighted collar on each end of the vial. Seventh, make a sling of masking tape attached to each side of each of the water baths to hold the pipettes out of the water during an equilibration period of 7 min. Vials 1, 2, and 3 should rest in the roomtemperature water bath and vials 4, 5, and 6 should rest in the 10 degree C water bath. Eighth, after the equilibration period of 7 min, immerse all six respirometers entirely in their water baths. Water will enter the pipettes for a short distance and then stop. If the water continues to move into the pipette, check for leaks in the respirometer. Work swiftly and arrange the pipettes so that they can be read through the water at the beginning of the experiment. They should not be shifted during the experiment. Hands should be kept out of the water bath after the experiment has started. Make sure that a constant temperature is maintained. Ninth, allow the respirometers to equilibrate for 3 more min and then record, to the nearest 0.01 mL, the initial position of water in each pipette. Check the temperature in both baths and record in Table 5.1. Every 5 min for 20 min, take readings of the water¶s position in each pipette. Record the data in Table 5.1.
Results The main result of this lab was that all data points towards the fact that germinating peas absorb more oxygen when kept at room temperature than at 10 degrees Celsius.
Table 1 Data: Beads Alone Temp. (C ) Time (Min) Initial 0 0-5 0-10 0-15 0-20 Initial 0 0-5 0-10 0-15 0-20 Germinating Peas Dry Peas and Beads Reading at Time X 9 1.4 .6 1.4 .6 8.2 7.5 7.3 7.3 9.5 1.2 1.5 1.2 .9 9.5 9.5 9.5 9.5 .8 .7 .2 0 Diff.*
Reading Reading at Time Diff.* Diff.* at Time x x 9.3 9.3 9.3 9.3 9.3 9.3 9.3 9.3 9.3 9.3 7 5.6 5 3.6 3 8.7 7.5 6 4.8 3.9
Although confusing, these results show that germinating peas absorb more oxygen than the dry peas at both temperatures. This also shows how the germinated peas absorbed more oxygen at room temperature than at 10 degrees Celsius. Also, the dead glass beads stayed at a constant rate of zero during the experiment. Figure 2:
10 9 8 7 10C Beads 6 5 4 3 2 1 0 Initial 5 Minutes 10 Minutes 15 Minutes 20 Minutes 10C Germ. 10C Non-Germ. 25C Beads 25C Germ. 25C Non-Germ.
(10C Beads and 25C Beads are in the same location all the way through)
Discussion Our results, seen in Figures 1 and 2, fully uphold our hypothesis (At a warmer temperature, more oxygen will be absorbed with the germinating peas than would be in cooler conditions). The data that best upholds our hypothesis is found in Figure 1, where you can see that the amount of oxygen absorbed is higher with the warmer temperature than the amount absorbed in the cooler temperature.
Conclusion After reviewing the results of this lab, I have learned that at a warmer temperature more oxygen will be absorbed with the germinating peas than would be at a cooler temperature, and that germinating peas absorb more oxygen than dry peas in both conditions.
Cellular Respiration Lab Manual
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