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BISC110: Series 3 Experiment 9 Hill Reaction: Difference between revisions
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BISC110: Series 3 Experiment 9 Hill Reaction (view source)
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4. Measurement of Hill Reaction Rates: For each reaction tube, do the following in turn and work as rapidly as possible and in dim light. Mix '''thylakoid suspension''' gently by flicking bottom of tube. | 4. Measurement of Hill Reaction Rates: For each reaction tube, do the following in turn and work as rapidly as possible and in dim light. Mix '''thylakoid suspension''' gently by flicking bottom of tube. | ||
5. Immediately remove 50µL of '''thylakoid suspension''' and add to a reaction tube containing 5mL of '''reaction | 5. Immediately remove 50µL of '''thylakoid suspension''' and add to a reaction tube containing 5mL of '''reaction mixture''', mix gently by inverting once and place tube in the Spec20 to obtain a "dark reading" with the spectrophotometer set at 580nm. This is the time = zero seconds absorbance reading. Take care not to shield the tube from the light with your hand. | ||
6. Take absorbance readings every 15s (10s illumination + 5s reading) for 90s (7 readings). Record absorbance readings and times in your lab notebook. Repeat the experiment, i.e., do several 90s runs, until you obtain the desired reference rate of DCPIP reduction. Then repeat the experiment, to see if you can reproduce your data consistently. | 6. Take absorbance readings every 15s (10s illumination + 5s reading) for 90s (7 readings). Record absorbance readings and times in your lab notebook. Repeat the experiment, i.e., do several 90s runs, until you obtain the desired reference rate of DCPIP reduction. Then repeat the experiment, to see if you can reproduce your data consistently. | ||
