Homework Answers
Actual substrate concentration:
Experiment #1. 1.5 mM * 1.5 mL/1.5 mL = 1.5 mM
Experiment #2. 1.0 mM * 1.0 mL/1.5 mL = 0.667 mM
Experiment #3. 0.5 mM * 0.5 mL/1.5 mL = 0.167 mM
Experiment #4. 0.25 mM * 0.25 mL/1.5 mL = 0.042 mM
Experiment #5. 1.125 mM * 0.12 mL/1.5 mL = 0.01 mM
Experiment #6. 0.06 mM * 0.06 mL/1.5 mL = 2.4*10-3 mM
Experiment #7. 0.03 mM * 0.03 mL/1.5 mL = 6*10-4 mM
Experiment #8. 0.015 mM * 0.015 mL/1.5 mL = 1.5*10-4 mM
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Experiment # Starting Substrate Concentration Actual Substrate Concentration Substrate Added (ml) Buffer Added (ml) 1.5 mm...
Table 2: Amounts of buffer, substrate, and enzyme added for lactase assay Tube 0.1 M 200...
Table 2: Amounts of buffer, substrate, and enzyme added for lactase assay Tube 0.1 M 200 uM Lactase Total Abs ONP PO4 ONPG added (420 nm) Formed (mL) (mL) (mL) (mL) (umol) 4.00 0.00 6.0 5.50 0.00 0.50 6.0 .120 5.25 0.50 6.0 -127 5.00 0.50 6.0 4.50 0.50 6.0 0176 -186 3.50 0.50 6.0 221 1.50 0.50 6.0 396 1. Calculate moles of ONP in Table 1. Show your work for any tube other than Tube 1 and 2....
Figure 8: ONP Formation v. Enzyme Conc. Enzyme Abs @ concentration 420 nm (units/ml) 0.2 0.372...
Figure 8: ONP Formation v. Enzyme Conc. Enzyme Abs @ concentration 420 nm (units/ml) 0.2 0.372 0.4 0.648 0.6 0.923 0.8 0.004 1.0 1.572 Examine Figure 8. In this experiment, all five samples were supposed to be subjected to identical conditions (substrate concentration, temperature, reaction time) except for the enzyme concentration, which is indicated in the table. Which of the following procedural errors could account for the unexpected result for the 0.8 units/ml enzyme concentration? This tube was left in...
In an 'Enzyme Kinetics' experiment, the following calibration data, product (p-nitrophenol, PNP) concentration (M) versus UV...
In an 'Enzyme Kinetics' experiment, the following calibration data, product (p-nitrophenol, PNP) concentration (M) versus UV Absorbance 405 nm, was obtained If the UV absorbance 405 nm within the 1.5 mL-cuvette was measured as 0.144 in one of the trials during the experiment, determine the amount of pNP (in mg) produced. Assume total volume of reaction mixture stays at 1.5 mL at all times. (Round up to two decimal places) Molecular Weight of PNP: 139.11 g/mol PNP Concentration UV Absorbancem...
2. Design a PCR experiment to amplify a sequence of interest. Use the following reagent concentrations...
2. Design a PCR experiment to amplify a sequence of interest. Use the following reagent concentrations for calculating your cocktail. Fill in the chart below. A. Fill in the chart below with the volumes that would be added to each tube. Note the final volume of the reaction. Show your calculations for full credit. REAGENT Final Concentration (50 ul Reaction) 1x Test Reaction (+) Control Reaction 10x Reaction Buffer ML dNTPs (15 mm) 200 UM 0.2 UM ul 0.2 um...
CALCULATIONS [2 points each] You are asked to perform a PCR experiment in the lab. However, in th...
CALCULATIONS [2 points each] You are asked to perform a PCR experiment in the lab. However, in this experiment you must add each ingredient separately (unlike the "master-mix" we used in our own experiment). In this experiment, each PCR reaction tube contained the following ingredients added with a micropipetor... 10 HL of 5x Buffer 6 μ1 of 25 mM MgCl2 solution μL of 10 mM dNTP mix 1.5 μL oligonucleotide primer mix 0.5 HL Taq DNA Polymerase stock solution 28...
Calculate initial concentration of Fe+3 for tubes 1-3. Show your work. Procedure A. Determination of B...
Calculate initial concentration of Fe+3 for tubes 1-3. Show your
work.
Procedure A. Determination of B for Beer's Law 1. Using a buret, add 4.00 mL of 0.0025 M Fe(NO3)s (which is in 0.1 M HNOs) to a 100- mL volumetric flask. Add enough deionized water to bring the total volume to the mark on the neck of the flask. Stopper and shake the flask. Label this flask “Diluted Fe.” spectrophotometer tubes (cuvettes), they are too small to use at...
3. Use the below tables to complete lab calculations on the worksheet on pages 2-6 of...
3. Use the below tables to complete lab calculations on the worksheet on pages 2-6 of this document. You will submit the worksheet via dropbox on Canvas by you assigned lab time the week of March 30th through April 3rd. A document with sample calculations of different concentrations is provided to you on canvas. Enzyme Kinetics Lab Buffer volume (mL) Enzyme Volume (ml) Substrate Volume (ml) TOTAL VOLUME (mL) 0.04 0.5 1.5 0.04 0.25 1.5 0.04 0.1 1.5 0.04 0.05...
my questions are the following: A) what is the concentration of the stock solution of LDH?...
my questions are the following:
A) what is the concentration of the stock solution of LDH?
B) Based upon Part B of the protocol, show your calculations to
make the 250ug/ml LDH solution.
C) Based upon Part B of the protocol, show your calculations to
make the 25ug/ml LDH solution.
You will be given 60 ul of LDH stock solution. You need to determine the protein concentration of this stock solution and then perform a serial dilution to end up...
4. (a) Determine the experimental concentration of protein in the unknown solution in mg/mL using the...
4. (a) Determine the experimental concentration of protein in the unknown solution in mg/mL using the Part II Data Table and the equation on page 2 for the absorbance assay at 280 nm. Hint: Convert the molar concentration (M or moles/liter) of BSA from absorbance assay at 280 nm method to mg/mL, assume that the molecular weight of BSA = 66.5 kDa [7 points 6500 g/mol Post-lab 10 Report Form: Determination the Concentration of a Protein You must show your...
Procedure: 1. 10 Test tubes were labelled (0C,0E,15C,15E,30C,30E,45C,45E,60C, and 60E.) 2. 2 mL cathecol-buffer at pH...
Procedure:
1. 10 Test tubes were labelled
(0C,0E,15C,15E,30C,30E,45C,45E,60C, and 60E.)
2. 2 mL cathecol-buffer at pH 6 was added into each test
tubes.
3. 3.5 mL of water was added into all of the C tubes
(0C,15C,30C,45C, and 60C.)
4. 3 mL of water was added into the remaining tubes.
5.The two tubes were put into an ice bath.
6. 0.5 mL of enzyme polyphenoloxidase was added into tube E when
the mixture had reached certain T (O°C)
7.The transmittance...
Table 2: Amounts of buffer, substrate, and enzyme added for lactase assay Tube 0.1 M 200 uM Lactase Total Abs ONP PO4 ONPG added (420 nm) Formed (mL) (mL) (mL) (mL) (umol) 4.00 0.00 6.0 5.50 0.00 0.50 6.0 .120 5.25 0.50 6.0 -127 5.00 0.50 6.0 4.50 0.50 6.0 0176 -186 3.50 0.50 6.0 221 1.50 0.50 6.0 396 1. Calculate moles of ONP in Table 1. Show your work for any tube other than Tube 1 and 2....
Figure 8: ONP Formation v. Enzyme Conc. Enzyme Abs @ concentration 420 nm (units/ml) 0.2 0.372 0.4 0.648 0.6 0.923 0.8 0.004 1.0 1.572 Examine Figure 8. In this experiment, all five samples were supposed to be subjected to identical conditions (substrate concentration, temperature, reaction time) except for the enzyme concentration, which is indicated in the table. Which of the following procedural errors could account for the unexpected result for the 0.8 units/ml enzyme concentration? This tube was left in...
In an 'Enzyme Kinetics' experiment, the following calibration data, product (p-nitrophenol, PNP) concentration (M) versus UV Absorbance 405 nm, was obtained If the UV absorbance 405 nm within the 1.5 mL-cuvette was measured as 0.144 in one of the trials during the experiment, determine the amount of pNP (in mg) produced. Assume total volume of reaction mixture stays at 1.5 mL at all times. (Round up to two decimal places) Molecular Weight of PNP: 139.11 g/mol PNP Concentration UV Absorbancem...
2. Design a PCR experiment to amplify a sequence of interest. Use the following reagent concentrations for calculating your cocktail. Fill in the chart below. A. Fill in the chart below with the volumes that would be added to each tube. Note the final volume of the reaction. Show your calculations for full credit. REAGENT Final Concentration (50 ul Reaction) 1x Test Reaction (+) Control Reaction 10x Reaction Buffer ML dNTPs (15 mm) 200 UM 0.2 UM ul 0.2 um...
CALCULATIONS [2 points each] You are asked to perform a PCR experiment in the lab. However, in this experiment you must add each ingredient separately (unlike the "master-mix" we used in our own experiment). In this experiment, each PCR reaction tube contained the following ingredients added with a micropipetor... 10 HL of 5x Buffer 6 μ1 of 25 mM MgCl2 solution μL of 10 mM dNTP mix 1.5 μL oligonucleotide primer mix 0.5 HL Taq DNA Polymerase stock solution 28...
Calculate initial concentration of Fe+3 for tubes 1-3. Show your
work.
Procedure A. Determination of B for Beer's Law 1. Using a buret, add 4.00 mL of 0.0025 M Fe(NO3)s (which is in 0.1 M HNOs) to a 100- mL volumetric flask. Add enough deionized water to bring the total volume to the mark on the neck of the flask. Stopper and shake the flask. Label this flask “Diluted Fe.” spectrophotometer tubes (cuvettes), they are too small to use at...
3. Use the below tables to complete lab calculations on the worksheet on pages 2-6 of this document. You will submit the worksheet via dropbox on Canvas by you assigned lab time the week of March 30th through April 3rd. A document with sample calculations of different concentrations is provided to you on canvas. Enzyme Kinetics Lab Buffer volume (mL) Enzyme Volume (ml) Substrate Volume (ml) TOTAL VOLUME (mL) 0.04 0.5 1.5 0.04 0.25 1.5 0.04 0.1 1.5 0.04 0.05...
my questions are the following:
A) what is the concentration of the stock solution of LDH?
B) Based upon Part B of the protocol, show your calculations to
make the 250ug/ml LDH solution.
C) Based upon Part B of the protocol, show your calculations to
make the 25ug/ml LDH solution.
You will be given 60 ul of LDH stock solution. You need to determine the protein concentration of this stock solution and then perform a serial dilution to end up...
4. (a) Determine the experimental concentration of protein in the unknown solution in mg/mL using the Part II Data Table and the equation on page 2 for the absorbance assay at 280 nm. Hint: Convert the molar concentration (M or moles/liter) of BSA from absorbance assay at 280 nm method to mg/mL, assume that the molecular weight of BSA = 66.5 kDa [7 points 6500 g/mol Post-lab 10 Report Form: Determination the Concentration of a Protein You must show your...
Procedure:
1. 10 Test tubes were labelled
(0C,0E,15C,15E,30C,30E,45C,45E,60C, and 60E.)
2. 2 mL cathecol-buffer at pH 6 was added into each test
tubes.
3. 3.5 mL of water was added into all of the C tubes
(0C,15C,30C,45C, and 60C.)
4. 3 mL of water was added into the remaining tubes.
5.The two tubes were put into an ice bath.
6. 0.5 mL of enzyme polyphenoloxidase was added into tube E when
the mixture had reached certain T (O°C)
7.The transmittance...
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