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3520
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Chemistry
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Feb 20, 2024
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Care and use of a Pipette and Practice Exercise
BIOC 3520
Abstract
This experiment was conducted to help students perfect their use of the micropipette and pipette, along with learning the proper care techniques for pipettes. The problem being addressed in this experiment is inaccurate pipette measurements and incorrect pipetting techniques. In this experiment, it was founded that if the pipette is consistently used correctly, meaning all the steps were followed, accurate and precise results can be
obtained each time unless there are other factors weighing in such as malfunctioning pipettes. The results from this experiment indicate that overall, the pipette is effective in measuring small volume amounts, and if used correctly, a percent error of 0% can be obtained. After completing the lab, it is better understood why individuals get inaccurate results and measurements and how this can be fixed to yield a low or 0% percent error.
Introduction
Pipettes are used to transfer specific liquid volumes into another container. In this experiment, the use of the pipette was practiced by making precise measurements. The
micropipette on the other hand was used to transfer small liquid volumes to other containers. The micropipette has a volume range that can vary depending on each micropipette. P20 is the small volume pipette ranging from 2-20ul. P200 is the mid-
range pipette ranging from 20-200ul. Lastly, P1000 is the large range micropipette ranging from 100-1000ul. When using pipettes, consistency is important to make sure you obtain accurate results. In this experiment, a certain volume amount was obtained 3
times and the mass, percent error, and standard deviation were all calculated to determine the accuracy of the pipette use which is shown in data table 1. In this experiment, if the pipette is used correctly, then the measurements will be precise and accurate.
Methods and Materials
Water
Pipette
Micropipette
Pipette tips
Fine balance
Microcentrifuge tube
Flask
Napkin (filter paper)
Laemmeli Buffer
Measuring accuracy of the micropipette
1.
First, after putting on your lab coats, obtain microcentrifuge tubes from the TA.
2.
Next, tare the scale and wait for it to show “0.00”. Then, measure the mass of your empty microcentrifuge tube then record it in your lab manual.
3.
After this, grab your pipette and plunge it into the correctly sized tip. Then, press the plunger to the first stop and place it into the water. Making sure the pipette tip
is still in the water, slowly release the plunger to obtain the liquid.
4.
After the correct volume is in the pipette, place the tip of the pipette into your empty microcentrifuge tubes and press the plunger to the first stop, then to the second stop to get any leftover liquids and close the microcentrifuge tube and expell the tip into your trash bag.
5.
Next, obtain the new mass of the microcentrifuge and record it in your lab manual.
6.
Using a new tip each time, repeat steps 2-5 3x for each of the volume amounts in
the table.
Results
Table 1
Micropipette
Volume
Expected
Weight
Actual
Weight
% error
Averag
e Error
Standard
Deviation
P20 (high volume)
20ul
0.02g
0.02g
0%
20ul
0.02g
0.03g
50%
20ul
0.02g
0.02g
0%
16.6%
0.00005
P200 (low volume)
30ul
0.03g
0.06g
50%
30ul
0.03g
0.03g
0%
30ul
0.03g
0.03g
0%
16.6%
0.0009
P200 (high volume)
180ul
0.180g
0.19g
5%
180ul
0.180g
0.18g
0%
180ul
0.180g
0.18g
0%
1.67%
0.005
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P1000 (low volume)
250ul
.250g
0.250g
0%
250ul
.250g
0.260g
4%
250ul
.250g
0.250g
0%
1.3%
2.30
P1000 (high volume)
850ul
.850g
0.850g
0%
850ul
.850g
0.850g
0%
850ul
.850g
0.850g
0%
0%
0
Questions from the manual
How does the percent error vary from trial to trial? Does your technique improve resulting in a smaller percent error with each trial? Explain why or why not.
For the most part, from trial to trial, the percent error decreased. This could mean our technique improved. For example, in our trials for the P200 high volume, we got 5%, 0%, and 0% respectively. It’s possible we withdrew the liquid too fast which would contribute to a percent error of 5%, but our technique improved which is why a percent error of 0% shows in the next trials.
Did the pipette volume affect your % error? Which pipette volume did you find the most difficult? What might be the reason for this?
Surprisingly, the pipette volume I found the most difficult was for P200 high volume and low volume. This volume had one of the highest percent errors out of all the trials. I’m not sure why though because I thought that the more volume, the more the accuracy
will go down. So, I was expecting P1000 to give me the most trouble.
Discussion
In this experiment, the results obtained were expected. Because this lab was for practicing the use of the pipette and micropipette, expecting perfect results was not realistic. The results in Table 1 were accurate for the most part, but in trials where there was a percent error, it could mean multiple things. It could mean the liquid was withdrew
too fast, or the pipette was not set correctly, or maybe the tip was not fully in the water which would lead to inaccuracy. Conclusion
Overall, in this experiment, the use a micropipette and pipette were effectively learned. It was learned that it is crucial to withdraw liquids slowly because doing it too fast can lead to an inaccurate volume. Another important tip that should be taken from this lab is determine which pipetting method works best for you, the reverse method, or the forward method. The importance of correctly setting the pipette was also learned. In the future, keeping these tips in mind will lead to results and measurements which are accurate. Another
References
N/A, only the lab manual was used.
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