Apparatus: Materials and Methods P1000 Gilson and blue gilson tips 3 disposable cuvettes Spectrophotometer parafilm stop watch tissue Chemicals: These chemicals are toxic, gloves and eye protection must be worn. All spills must be reported to the lecturer or a demonstrator and cleaned up immediately. Students must wash their hands before the laboratory is left. 50 mM sodium phosphate buffer, pH 7.5 (20ml) Distilled water 2 mM 2-nitrophenyl-ß-D-galactopyranoside (ONP-Gal) (5ml) 2 mM 2-nitrophenyl-ß-glucopyranoside (ONP-Glu) (5ml) 2 units.ml-¹ ß-galactosidase (from E. coli) (10ml) Method: 1. Check that the spectrophotometer is on and set at 440 nm. If it is not set up correctly ASK for help. 2. Fill the first cuvotto with f
Apparatus: Materials and Methods P1000 Gilson and blue gilson tips 3 disposable cuvettes Spectrophotometer parafilm stop watch tissue Chemicals: These chemicals are toxic, gloves and eye protection must be worn. All spills must be reported to the lecturer or a demonstrator and cleaned up immediately. Students must wash their hands before the laboratory is left. 50 mM sodium phosphate buffer, pH 7.5 (20ml) Distilled water 2 mM 2-nitrophenyl-ß-D-galactopyranoside (ONP-Gal) (5ml) 2 mM 2-nitrophenyl-ß-glucopyranoside (ONP-Glu) (5ml) 2 units.ml-¹ ß-galactosidase (from E. coli) (10ml) Method: 1. Check that the spectrophotometer is on and set at 440 nm. If it is not set up correctly ASK for help. 2. Fill the first cuvotto with f
Chemistry
10th Edition
ISBN:9781305957404
Author:Steven S. Zumdahl, Susan A. Zumdahl, Donald J. DeCoste
Publisher:Steven S. Zumdahl, Susan A. Zumdahl, Donald J. DeCoste
Chapter1: Chemical Foundations
Section: Chapter Questions
Problem 1RQ: Define and explain the differences between the following terms. a. law and theory b. theory and...
Related questions
Question
![Apparatus:
●●●●
●●●
P1000 Gilson and blue gilson tips
3 disposable cuvettes
Spectrophotometer
parafilm
stop watch
tissue
Materials and Methods
Chemicals:
These chemicals are toxic, gloves and eye protection must be worn. All spills must be reported
to the lecturer or a demonstrator and cleaned up immediately. Students must wash their hands
before the laboratory is left.
50 mM sodium phosphate buffer, pH 7.5 (20ml)
Distilled water
2 mM 2-nitrophenyl-ß-D-galactopyranoside (ONP-Gal) (5ml)
2 mM 2-nitrophenyl-ß-glucopyranoside (ONP-Glu) (5ml)
2 units.ml-¹ B-galactosidase (from E. coli) (10ml)
Method:
1. Check that the spectrophotometer is on and set at 440 nm. If it is not set up correctly ASK for
help.
3. In the second cuvette add:
beslist elven
2. Fill the first cuvette with 3 ml distilled water and zero the spectrophotometer by placing the
cuvette in the machine in the correct orientation and pressing the BLUE button. If you follow the
instructions below you will not need to repeat this.
1 ml 50 mM sodium phosphate buffer, pH 7.5
1 ml Distilled water
0.5 ml 2-nitrophenyl-ß-D-galactopyranoside (ONP-Gal)
<pause here before moving to next step>
add 0.5ml ß-galactosidase to the cuvette WHEN YOU ARE READY TO START
THE CLOCK
Mix by covering the top of the cuvette with parafilm, ensure the outside of the cuvette is dry and
place in the correct orientation (bevelled sides to the left and right) in the spectrophotometer.
Press the GREEN button to start measuring the absorbance.
4. Take your first absorbance reading at 30 seconds and continue taking readings every 30
seconds for 5 minutes, record the reading in the form of a table (for table, see next page).
5. REPEAT the experiment starting at step 3.
6. Calculate the average from the 2 experiments for each time point. Plot your data on a graph
and calculate the rate (absorbance units minute).
Rate of the reaction based on your plotted values:](/v2/_next/image?url=https%3A%2F%2Fcontent.bartleby.com%2Fqna-images%2Fquestion%2Fbbba6467-9518-4216-af68-86921dbd3664%2F92c44f82-5f9e-4cfb-8ff5-c854d322a3a1%2F52inr5_processed.jpeg&w=3840&q=75)
Transcribed Image Text:Apparatus:
●●●●
●●●
P1000 Gilson and blue gilson tips
3 disposable cuvettes
Spectrophotometer
parafilm
stop watch
tissue
Materials and Methods
Chemicals:
These chemicals are toxic, gloves and eye protection must be worn. All spills must be reported
to the lecturer or a demonstrator and cleaned up immediately. Students must wash their hands
before the laboratory is left.
50 mM sodium phosphate buffer, pH 7.5 (20ml)
Distilled water
2 mM 2-nitrophenyl-ß-D-galactopyranoside (ONP-Gal) (5ml)
2 mM 2-nitrophenyl-ß-glucopyranoside (ONP-Glu) (5ml)
2 units.ml-¹ B-galactosidase (from E. coli) (10ml)
Method:
1. Check that the spectrophotometer is on and set at 440 nm. If it is not set up correctly ASK for
help.
3. In the second cuvette add:
beslist elven
2. Fill the first cuvette with 3 ml distilled water and zero the spectrophotometer by placing the
cuvette in the machine in the correct orientation and pressing the BLUE button. If you follow the
instructions below you will not need to repeat this.
1 ml 50 mM sodium phosphate buffer, pH 7.5
1 ml Distilled water
0.5 ml 2-nitrophenyl-ß-D-galactopyranoside (ONP-Gal)
<pause here before moving to next step>
add 0.5ml ß-galactosidase to the cuvette WHEN YOU ARE READY TO START
THE CLOCK
Mix by covering the top of the cuvette with parafilm, ensure the outside of the cuvette is dry and
place in the correct orientation (bevelled sides to the left and right) in the spectrophotometer.
Press the GREEN button to start measuring the absorbance.
4. Take your first absorbance reading at 30 seconds and continue taking readings every 30
seconds for 5 minutes, record the reading in the form of a table (for table, see next page).
5. REPEAT the experiment starting at step 3.
6. Calculate the average from the 2 experiments for each time point. Plot your data on a graph
and calculate the rate (absorbance units minute).
Rate of the reaction based on your plotted values:
![Experiment 1
Time (Minutes) Absorbance level (AU)
0.12
0.181
0.274
0.364
0.451
0.533
0.606
0.675
0.734
0.786
0.5
1
1.5
2
2.5
3
3.5
4
4.5
5](/v2/_next/image?url=https%3A%2F%2Fcontent.bartleby.com%2Fqna-images%2Fquestion%2Fbbba6467-9518-4216-af68-86921dbd3664%2F92c44f82-5f9e-4cfb-8ff5-c854d322a3a1%2Festoq4_processed.jpeg&w=3840&q=75)
Transcribed Image Text:Experiment 1
Time (Minutes) Absorbance level (AU)
0.12
0.181
0.274
0.364
0.451
0.533
0.606
0.675
0.734
0.786
0.5
1
1.5
2
2.5
3
3.5
4
4.5
5
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