Flow Chart For Determining Enzyme
Flow Chart For Determining Enzyme
Flow Chart For Determining Enzyme
BIOC 2069
PRACTICAL # 1
Instrumentation in the Laboratory
Student Learning Outcomes
At the end of this lab, you should be able to:
MEASURING pH
The pH meter is a simple machine used to measure pH. It has two main components - an
electrode which is usually on a stand and the base immersed in buffer or distilled water and a
dashboard which has a display.
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BIOC2069
Lab 1 Instrumentation
Semester I 2016/2017
Procedure
You will be provided with 0.5M HCl, 0.25 M HCl and 0.1M HCl solutions on the centre bench.
1. Place the electrode into your sample (b) (0.25M NaOH) from above.
2. Using a Pasteur pipette in a drop wise manner directly introduce 0.5M HCl until the pH nears
6.8-7.2. Ensure that after each addition of acid, the solution is well vortexed or swirled before
taking the final pH reading. It is not necessary to rinse or remove the electrode from the
solution during the procedure.
3. Once the pH is around 6.8-7.2, use the 0.25M HCl to adjust the pH to around 6.
4. Use the 0.1 M HCl to further adjust the pH to 5.
5. Adjust the volume to 10mL with distilled water. If you pass the 10mL mark, measure and
record the final volume.
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BIOC2069
Lab 1 Instrumentation
Semester I 2016/2017
Tube number
Solution 1
Solution 2
Solution 3
Solution 4
4 L
4 L
2 L
4 L
4 L
2 L
3 L
3 L
2 L
2 L
Total volume
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BIOC2069
Lab 1 Instrumentation
Semester I 2016/2017
8. A total of 10 L of reagents was added to each reaction tube. To check the measurements were
accurate, set the pipette to 10 L and very carefully withdraw solution from each tube.
a. Is the tip just filled? or
b. Is a small volume of fluid left in tube? or
c. After extracting all fluid, is an air space left in the tip end? (The air can be displaced and actual
volume determined simply by rotating, volume adjustment to push fluid to very end of tip. Then,
read the volume directly).
b) Large-Volume Micropipettor
This exercise simulates a bacterial transformation or plasmid preparation, for which a 100-1000
L micropipettor is used. It is far easier to mismeasure when using a large-volume micropipettor.
If the plunger is not released slowly, an air bubble may form or solution may be drawn into
piston.
Tube number
Solution 1
Solution 2
Solution 3
Solution 4
100 L
200 L
150 L
550 L
150 L
250 L
350 L
250 L
Total volume
3. Set the micropipettor to add appropriate volumes of Solutions 1-4 to reaction tubes D and E.
Follow the same procedure as for the small-volume micropipettor exercise.
4. A total of 1000 L of reactants was added to each tube. To check that measurements were
accurate, set the micropipettor to 1000 L and carefully withdraw solution from each tube.
5. a. Is the tip just filled? or
b. Is a small volume of fluid left in tube? or
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BIOC2069
Lab 1 Instrumentation
Semester I 2016/2017
c. After extracting all fluid, is an air space left in the tip end? (The air can be displaced and actual
volume determined simply by rotating volume adjustment to push fluid to very end of tip. Then,
read the volume directly.)
the origin??) The absorbance value of the unknown is read on the spectrophotometer and should
fall within the line of your curve. Once you have determined the amount of your unknown you
can work back to determine its concentration. For your lab submissions, all calibration curves
must be digitally drawn with the equation written on the same page next to the line (use Excel).
The equation can be used to determine the amounts in your unknowns.
MATERIALS
Reagent A: 2%Na2CO3 on 0.1M NaOH
Reagent B: 0.5% CuSO4.5H2O in 1% sodium citrate
Reagent C: Mix just before using. Combine Reagent A and Reagent B in a 50:1 mL ratio
Diluted Folin Cicocalteaus Reagent (1:1 with water) this has been provided with you