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BISC110/F12: Series 3 Experiment 9 Hill Reaction: Difference between revisions
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BISC110/F12: Series 3 Experiment 9 Hill Reaction (view source)
Revision as of 11:02, 10 May 2012
, 10 May 2012→Procedure
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#When you have set your light intensity to an even 80 µmol photons m-2 s-1 illumination of a tube that is held steadily in the tube holder apparatus you are ready to start your first trial reaction. | #When you have set your light intensity to an even 80 µmol photons m-2 s-1 illumination of a tube that is held steadily in the tube holder apparatus you are ready to start your first trial reaction. | ||
#Immediately remove 50 µL of the undiluted stock thylakoid suspension and add it to a reaction tube containing 5 mL of reaction mixture, mix gently by inverting once and place tube in the Spec20 to obtain a "dark reading" with the spectrophotometer set at 580 nm. This is the time = zero seconds absorbance reading. Expose the thylakoids to the light and take care not to shield the tube from the light with your hand. | #Immediately remove 50 µL of the undiluted stock thylakoid suspension and add it to a reaction tube containing 5 mL of reaction mixture, mix gently by inverting once and place tube in the Spec20 to obtain a "dark reading" with the spectrophotometer set at 580 nm. This is the time = zero seconds absorbance reading. Expose the thylakoids to the light and take care not to shield the tube from the light with your hand. | ||
#Take absorbance readings every 15 s (10 s illumination + 5 s reading) for 90 s (7 readings). Record absorbance readings and times in your lab notebook. Repeat the experiment at a light intensity | #Take absorbance readings every 15 s (10 s illumination + 5 s reading) for 90 s (7 readings). Record absorbance readings and times in your lab notebook. Repeat the experiment at a higher light intensity (intensity is your choice but should be higher than 120 µmol photons m-2 s-1, using undiluted thylakoids). Repeat the experiment again using the 1/2 dilution of thylakoids at 80 µmol photons m-2 s-1 and at your higher light intensity. Run the lower light intensity for 2 minutes rather than for 90 seconds. <BR> | ||
Compare your different trials to the following general standards:<BR> | Compare your different trials to the following general standards:<BR> | ||
