C. Spectrophotometric Measurements of the Hill Reaction1.Place 15 mL of the 0.05-mg chlorophyll/ mL suspension in a test tube. Wrap thetube with aluminum foil and chill in an ice bucket. This is the source of unheatedchloroplasts.2. Place the remaining 10 mL of the 0.05-mg chlorophyll/mL suspension in a testtube and heat until boiling using an alcohol lamp. Heating inactivates thechloroplasts. Stand the heated mixture for 5 min. to allow the debris to settle. neeea3. Prepare the contents of each setup listed in Table 5.1 in clean test tubes. Pipetcontents in each test tube in the order given. For the "Unheated (Dark)" setup,immediately wrap the test tube with aluminum foil even before the first reading ismade.Table 5.1. Contents of tubes for spectrophotometric determination of the Hill reaction.PBSContentsHeatedchloroplastsUnheatedchloroplasts0.1% DPIP(to be addedimmediatelybefore the firstreading)Heated3.51.0Volume Added (mL)BlanksUnheated3.51.0Heated2.51.01.0Experimental set-upsUnheated(Light)2.51.01.0Unheated(Dark)2.51.01.04. Your instructor will demonstrate the handling of a cuvette and operating aspectrophotometer. Set the spectrophotometer at 605 nm wavelength. Zero thespectrophotometer using the appropriate blank. Use the "Heated Blank" for"Heated" experimental tube, and use the "Unheated Blank" for both "Unheated(Light)”, and “Unheated (Dark)" experimental tubes.5. Add 1 mL 0.1% DPIP to experimental tubes "Heated", "Unheated (Light)", and"Unheated (Dark)" prior to the initial readings. DPIP must be added toexperimental tubes when about to be placed in spectrophomoter. Cover themouth of the test tube with a piece of aluminum foil and the thumb and invert thetest tube several times to mix its contents. Pour the solution in a clean cuvette. Donot delay reading if DPIP was already mixed.6. Take the initial readings of the experimental set-ups "Heated", "Unheated (Light)",and "Unheated (Dark)". For the "Unheated (Dark)" setup, immediately wrap thetest tube in aluminum foil after taking the initial reading. Measure the absorbanceagain after 30 min. Uncover only upon reading the absorbance.7. After the initial readings, place all the test tubes in a beaker placed in an ice bathand illuminate the test tubes with a 100-watt bulb placed 18 inches away. Measurethe absorbance of the "Heated" and the "Unheated (Light)" tubes at 5-min. intervalsup to 30 min. Mix the contents before reading.8. Record all the data as Table 5.1 in your laboratory notebook.

It is a typical technique for figuring out how many chemicals are present in a solution or how much…

C. Spectrophotometric Measurements of the Hill Reaction 1. Place 15 mL of the 0.05-mg chlorophyll/ mL suspension in a test tube. Wrap the tube with aluminum foil and chill in an ice bucket. This is the source of unheated chloroplasts. 2. Place the remaining 10 mL of the 0.05-mg chlorophyll/mL suspension in a test tube and heat until boiling using an alcohol lamp. Heating inactivates the chloroplasts. Stand the heated mixture for 5 min. to allow the debris to settle.

C. Spectrophotometric Measurements of the Hill Reaction 1. Place 15 mL of the 0.05-mg chlorophyll/ mL suspension in a test tube. Wrap the tube with aluminum foil and chill in an ice bucket. This is the source of unheated chloroplasts. 2. Place the remaining 10 mL of the 0.05-mg chlorophyll/mL suspension in a test tube and heat until boiling using an alcohol lamp. Heating inactivates the chloroplasts. Stand the heated mixture for 5 min. to allow the debris to settle.

Basic Clinical Laboratory Techniques 6E

6th Edition

ISBN:9781133893943

Author:ESTRIDGE

Publisher:ESTRIDGE

Chapter6: Basic Clinical Chemistry

Section6.6: Blood Cholesterol And Triglycerides

Problem 8RQ

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