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Experiment #1
Pipette Calibration
Introduction
Automatic adjustable pipettes have become an
indispensable tool in today's biochemistry laboratory.
They are highly advantageous due to their accuracy,
ease of use, and wide range of volume deliveries
available.
PIPETTE
Eppendorf 10
0.5-10
Eppendorf 100
10-100
Eppendorf 1000
100-1000
Operation
Each pipette is equipped with a comfortable handgrip, a
disposal button, a delivery button, and a grooved wheel for
volume delivery adjustment. The delivery button, when
pushed, is equipped with two "STOP" points, noticeable by the
feel of gentle resistance. The first STOP is to obtain the
Page: 2
PIPETTE
Eppendorf
10
Eppendorf
100
Eppendorf
1000
PIPETTE READING
Max.
reading
Max.
reading
Max.
reading
= 10.00
L
= 8.55
L
= 100.0
L
= 52.5
L
= 1000
L
= 525
L
Page: 3
Experiment
All adjustable pipettes should have their calibration checked
periodically. The easiest way to accomplish this task is to
pipette an indicated volume of a solution of known density,
weigh the amount transferred, and determine if the
measured volume corresponds to the indicated volume. The
density of distilled water at 25 o C is 0.997075 g/ml; an
DENSITY
(g/mL)
20
0.998234
21
0.998022
22
0.997801
23
0.997569
24
0.997327
25
0.997075
26
0.996814
27
0.996544
28
0.996264
Page: 4
Data Analysis
1. Using a computer program type in each of the
datasets as (cumulative weight - weight of empty
boat) versus (total volume added). Plot each dataset
(linear option) and fit each plot with a linear
regression. Record the slope of the plot, the
correlation coefficient and obtain an error estimate for
each dataset. Printouts of these plots should be
included in your lab notebook.
2. For all three datasets, convert the cumulative weights
into a table of weights per aliquot. Enter these data
into the spreadsheet and obtain appropriate statistics
for each set. Divide the mean weights by the density
(corrected for temperature) to obtain the mean
volumes delivered. Calculate the percent error for each
of the three volume pipettes.
In your laboratory notebook you should include the plots
prepared above and a table reporting slopes, and the mean,
median and standard deviation for each dataset, as
determined from your analysis. A second section of the table
should report these same data as obtained by your lab
partner. You should also comment on the accuracy and the
reproducibility of your pipetting, a comparison of your error
and your partner's and the correlation between the volumes
indicated on the pipettes and the actual volumes delivered.
Please note that your notebook will be scrutinized by your TA
and you should utilize this opportunity to practice writing for
future and more involved laboratory reports.