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For
the
experiment,
two
instruments were used in order to test the
accuracy and precision of the operators.
In order to test for the accuracy
of the operators, the red and blue
micropipettors with its white and blue
tip,
made
from
polypropylene,
respectively, were used to transfer 1000
mL of distilled water and the required
volumes of bromophenol blue (uL) to
the microcentrifuge tubes. One member
only did the transfer of the set volumes
of 0.5, 1.0, 1.5, 2.0 and 2.5 L of
Bromophenol blue and 1000 mL of
distilled water. Once the liquid has been
transferred to the microcentrifuge tube, it
was then vortexed to mix the contents of
the tube until the dye immersed in the
solution.
After the tubes have been
prepared, the spectrophotometer was
warmed up and set to a wavelengthmeasured 540nm. With distilled water
(blank solution), the spectrophotometer
was set to zero. Once calibrated, the
absorbance of the dye solutions was read
by the spectrophotometer starting from
the least concentrated to the most
concentrated.
In order to test for the precision
of operators, again, the red and blue
micropipettors, with its white and blue
tips, respectively, were utilized to
transfer the required volume of 2.5 L to
the microcentrifuge tubes. All the
members took part in this process as the
precision of each student in utilizing the
micropipettor
was
taken
into
consideration for this portion of the
experiment. Each member, using the
blue micropipettor with its blue tip
transferred
1000
mL
to
the
microcentrifuge tube, then afterwards, a
volume of 2.5 L of Bromophenol blue
was placed in each of the five tubes.
Once the solution has been prepared, the
tubes were then vortexed in order to mix
the contents of the tube and immerse the
dye into the solution, the tube was
placed to the microcentrifuge. After the
tubes have been prepared, the tubes,
together with the reagent blank was
placed in the spectrophotometer, which
read the absorbance of the dye solutions
starting from the least concentrated to
the most concentrated.
Sample Preparation
1000 mL of distilled water was
transferred to an empty microcentrifuge tube
using the blue micropipettor and 0.5 L of
Bromophenol blue was transferred to the
same microcentrifuge tube using the red
micropipettor. Once the tube has been
prepared, the tube was vortexed for 5
seconds to mix the contents of the solution.
The spectrophotometer, set to the
wavelength of 540 nm was calibrated and
set to zero using the blank solution. Once
calibrated, the absorbance of the tube
containing water and Bromophenol blue was
read.