In part II of the experiment, what is the initial concentration of glucose in th

Question

In part II of the experiment, what is the initial concentration of glucose in the first cuvette (after you mix glucose with assay mix, but before reaction changes the initial concentrations)? Report numeric answer in the units of molarity (M), but do not enter the units - numbers only.

or "blank" the Substrate Concentration six large test tubes from your drawer) and add water and 0.30 M glucose c. Velocity vs. Sub 1 the table helow. Mix the tubes thoroughly. using parafilm. table below. Mix the tubes thoroughly, using parafilm. xidase assay acconding to the mL glucose mL water tube 0.50 1.00 2.00 4.00 9.50 9.00 8.00 6.00 3.00 none Gars 10.00 2. Prepare six plastic assay cuvettes with 2.0 ml of glucose oxidase assay mix in each cuvette. Make sure to use the Part 2 glucose oxidase assay mirardenp Next, calibrate the spectrophotometer by using one of the cuvettes containing assay ero the spectrophotometer (see step 8 in Setting up the Spectrophotometer) as dy to 4. Now, do the following steps, being ready to start collecting data immediately after the addition of glucose a. Add 0.5 mL of glucose from large test tube #1 (step 1) into the first assay cuvette b b. Rapidly cover with Paralilm and invert twice to mix. c. Rapidly wipe cuvette sides with a Kimwipe, insert the cuvette into the spectrophotometer and press green start arrow to start data collection (you may need to push "Erase and Continue" if there is already data on the screen) d. Data will begin to appear. When data reaches 1 min push the red stop button. e. Go to "Analyze" at top tool bar. Select "Linear Fir f. Record slope on your lab report including units g. C the to ontinue with remaining cuvettes. Slope will appear automatically for the remaining cuvettes if you push "Erase and Continue" each time. Excel, to make a saturation plot. Also plot 1/velocity versus 1/substrate concentration in Excel to make a double-reciprocal plot. Your graphs should look something like the images below. 5. Plot Velocity (Abs / min) versus Substrate Concentration (final molarity of glucose) in 41

Explanation / Answer

Ans. The total constitution of reaction mixture in cuvette 1 is-

            0.50 mL of 0.30 M glucose + 9.50 mL water + 2.0 mL assay mix

# Total volume of reaction mixture = 0.50 mL + 9.50 mL + 2.0 mL = 12.0 mL

# Using           C1V1 (stock glucose solution) = C2V2 (Cuvette 1 reaction mixture)

            Or, 0.30M x 0.50 mL = C2 x 12.0 mL

            Or, C2 = (0.30M x 0.50 mL) / 12.0 mL

            Hence, C2 = 0.0125 M

Therefore, [Glucose] in the cuvette 1 reaction mixture = 0.0125 M

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