Question: Detection Methods For Biomolecules And Introduction To The Scientific Method 1 Now The Difficult Part: Write The Experimental Procedure. For Proper Experimental Design, A Lot Of Time Needed To Establish The Proportion Of Various Chemicals Used In Cach Test Condition So They Can Result In A Meaningful Outcome. Too Much Or Too Little Of A Certain Component …

Detection Methods for Biomolecules and Introduction to the Scientific Method 1 Now the difficult part: Write the experimental procedure. For proper experimental design, a lot of time needed to establish the proportion of various chemicals used in cach test condition so they can result in a meaningful outcome. Too much or too little of a certain component (starch or enzyme in this case) might result in an outcome that cannot be measured, or all samples will look more or less the same. To avoid those possibilities some guidelines are provided: Each sample will contain 3 ml of starch and 1 ml of amylase. (Are there any samples that will not have these components? 2. Incubate each component of the samples (starch and enzyme) separately at the conditions tested for 5 minutes before mixing. (Pour the starch into the tube with amylase after the five minutes of pre- incubation time.) 3. Once mixed, incubate them for exactly 20 minutes. 4. Prepare the controls. 5. Stop the reaction by adding 1 ml acid ethanol. (Why will this stop the reaction! 6. Add three drops of IKI to each of the samples, including the controls, mix. 7. Measure the outcome color intensity) using a spectrophotometer set at 670 nm. As demonstrated in the previous laboratory, color development and its intensity can serve as indicators of presence and relative amounts of the detected molecule. What represents a positive reaction for presence of starch in an IKI test? The intensity of this color will serve as a measurement of the activity of amylase: darker color indicates more starch is present, the enzyme was less efficient in the hydrolysis of starch: lighter color indicates the enzyme has higher efficiency in the hydrolysis of starch. Since the color intensity can vary, it is often dif- ficult to have quantitative value assigned to it without measuring equipment. In this experiment, a digital spectrophotometer will be used to quantify the color intensity of each reaction and indirectly, the activity of the enzyme reaction. The instructor will explain how the spectrophotometer works, and a brief outline is given in Appendix II. The procedure for use of the spectrophotometer: Turn on the spectrophotometer and allow it to warm up for 5 minutes. b. Verify correct wavelength of 670 nm. c. Set the machine to measure transmission by pressing the Trans! Abs button until the indicator light is on below the Trans label d. Calibrate the machine with the negative control (no starch + IKI-amber color solution). Wipe the tube clean with paper towel, open the cover of the sample compartment, and place the tube in it. Close the cover Press the 100% Trans/ Abs button once and wait for the machine to display 100% on the LCD screen. The machine is now calibrated and from this point on, only the tubes with the samples are sequen- tially placed in the sample compartment and no other calibrations or adjustments are needed. Remove the negative control from the compartment and proceed with the next step, f Place the positive control (starch that was not hydrolyzed) in the compartment and close the cover. Do not touch any buttons on the machine, simply read the measurement on the screen. After the mea. surement is recorded, remove the tube from the compartment. & Repeat step () for each of the samples in the experiment. Read and record the values of transmitted light