AN EXPERIMENT ON THE PRECISION AND ACCURACY OF USING THE MICROPIPETTE

Gyle Bennett Pambid¹, Park SangKeon¹, Kyla Nicole Ponce¹, & Charisse Mae Pondoc¹

¹Department of Biological Sciences, University of Santo Tomas

Abstract: The micropipette is an essential instrument in molecular biology laboratories. It is the perfect
tool in obtaining quantifications and statistical data. This experiment tests the precision and accuracy in
using the micropipette. Obtaining the raw data for testing the precision was done by weighing 50µL,
100µL, and 200µL of distilled water and was repeated three times for each volume, respectively. The
raw data for testing accuracy was obtained by weighing 75µL of distilled water, done by three different
persons. The recorded raw data were computed to determine the average weight, standard deviation
and percent error. The results show that both the standard deviation and standard error are low which
means that the results were evenly stable and the test was highly reliable, respectively. The data also
showed low variability which signifies great precision. The accuracy and precision of the samples can be
influenced by human error and various factors when handling and calibrating using the micropipette.
Strict compliance with the proper procedures when handling micropipettes should be observed and
understood to deliver excellent results of data.

Keywords: accuracy, micropipette, precision, biotechnology

INTRODUCTION and calculations with its calibrations will ensue

Biotechnology scopes different aspects after the experiment.
in the fields of both science and technology,
METHODS
being significant to molecular biology, and it
The experiment was done in two parts.
includes findings in areas of genomics,
The first part is testing the accuracy in using the
proteomics, DNA, and RNA isolation. One
micropipette. An analytical balance was used to
essential instrument for molecular biology
weigh 50µL, 100µL, and 200µL of distilled water
laboratories is micropipettes. Micropipettes
and the weight is recorded, up to 4 decimal
have a finer calibration than other instruments,
places. The procedure was done three times for
which varies from four types (p2, p20, p200,
each volume, respectively. The average weight
and p1000), making them effective in
and percent error were computed from the
distributing extreme volumes (microliters),
obtained data. The last part is testing the
ensuring both accuracy and precision.
precision in using the micropipette. The
Furthermore, quantifications and statistical data
analytical balance was used to weigh 75µL of
are often tackled in most lab experiments,
distilled water and the weight is recorded, up to
having micropipettes to be the perfect tool in
4 decimal places. This was done by three
obtaining these data. Familiarity with the use of
different persons. The average weight and
micropipette with the procedures, techniques,
standard deviation were computed from the
obtained data.

1
 2

As seen in table 1, small percent error

was computed from the acquisition of 50µL
with 2.87% compared to the 14.86% and 5.09%
FINDINGS accumulated from the samples with a volume of
The experiment aims for students to 100µL and 200µL, respectively, which is in
manipulate the micropipette properly. Table 1 contrast to observations stating that the
shows the weight of dH2O that students dispensing smaller volumes are more prone to
measured with micropipette. Using the average error compared to pipetting larger volumes.
of three trials, the percent error of experiment This is due to the surface tension at the tips of
measuring 50μL and 200μL was calculated near the micropipette, leaving some amounts of
5%, but the % error for the 100μL volume was sample to be discarded, thus strict compliance
14.86%, which means there is a possibility of with the procedure given as well as adept
mistake from measuring. techniques should be done to achieve
Table 2 calculates the precision of the maximum accuracy of samples. Consequently,
result which represents the standard deviation the data showed low variability thus signifies
of the given data values at 1.58 and the great precision within the values as well as the
standard error is 0.91. The low standard sample mean, which makes a great estimate for
the population mean.
Table 1. Testing Accuracy using the micropipette
Furthermore, accuracy and precision of
WEIGHT (mg)
VOLUME % error the samples can be influenced by various
Trial 1 Trial 2 Trial 3 Average factors such as: 1) environmental factors such
50 μL 46.7 49.8 48.8 48.4 2.87% as temperature, altitude and heat transfers; and
100 μL 132.1 100.1 111.2 114.5 14.86% 2) human errors involving techniques in
200 μL 222.1 203.2 203.1 209.5 5.09% micropipetting (Groot, 2018; Pushparaj, 2020).
Sample solutions that were subjected to higher
or lower temperatures deliver lower volumes
Table 2. Testing Precision using the micropipette compared to samples subjected to ambient
GROUP Standard
MEMBER
WEIGHT (mg)
Deviation
Standard Error conditions which is because of the expansion
1.Pambid, Gyle 46.7 46.7 46.7 and contraction of air volume confined which is
2.Park, Sang
49.8 49.8 49.8 also influenced by altitude given that it
Keon
3.Ponce, Kyla 48.8 48.8 48.8
decreases the air-cushion thus limiting the
4.Pondoc, acquisition of samples. Furthermore, heat
- -
Charisse -
transfers from the hand during prolonged
Overall Average 48.43 1.58 0.91
pipetting would drive warmth in the barrel of
the micropipette thus increasing the confined
deviation means that the result of volume of air and decreasing the accumulated
measurement data was evenly stable. Also, the volume of the sample (Pushparaj, 2020).
low standard error emphasized that the test Human errors involves various
was highly reliable. techniques in micropipetting such as: a) pre-
rinsing of pipette tips to an aliquot of the
DISCUSSION sample increases the trueness of the samples
collected given that capillary effects and air

temperature within the tip is being neutralized
(Pushparaj, 2020) thus pre-wetted tips deliver
accurate volumes of samples compared to dry
pipette tips, which delivers lower volume given
that pre-wetted tips have ambient humidity
which limits the amount of evaporation of the
sample (Joyce and Tyler, 1973); b) rhythm of
dispensing given that inconsistent aspiration
may cause the formation of aerosols and
contamination in the micropipette as well as
decrease the volume and efficiency of sample
volume (Pushparaj, 2020); c) the position of the
micropipette during calibration such that the
micropipette is at an angled and not in vertical
position will increase the amount of liquid to be
aspirated due to hydrostatic pressure (Almeida,
Batista and Felipe, 2013); d) the depth of
immersion given that it may increase the
sample volume, if too deep due to the
adherence of the sample at the tip thus may be
transferred along with the aliquot or may
decrease the sample volume, if near the surface
which let air to be aspirated, as well (Almeida,
Batista and Felipe, 2013, Pushparaj, 2020); and
the e) angle of dispensing which is at 45 degrees
with respect to the wall of the tube to prevent
the sample from being adhesive to the vent of
the tube (Ewald, 2015; Pushparaj, 2020).
CONCLUSION
Accuracy and precision can be affected
by human error and various factors when
handling and calibrating using the micropipette.
Trials show evenly measured data and a highly
reliable test result due to a low standard
deviation (Table 2) yet differs with its respective
percentage error, particularly with measuring
higher volumes (Table 1) which is a probable
human error in both handling and calibrating
the micropipette. Furthermore, strict
compliance with the said procedures when
3
handling micropipettes should be observed and
understood to deliver excellent results of data.
ACKNOWLEDGMENT
The authors would like to extend their
gratitude to the instructors of the BIO4212L
course, for providing collated data for this
formal report.
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doi:10.6026/9732063001600