Biotek Synergy LX, User Manual

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71 | Chapter 5 : Instrument Qualification Process
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For each Mean, calculate the Allowed Deviation using the Repeatability
specification for a 96-well plate of ±1.0% ±0.005 OD.
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The Standard Deviation for each set of readings should be less than the Allowed
Deviation.
Example: Readings of 1.950, 1.948, 1.955, 1.952, and 1.950 will result in a Mean of
1.951, and a Standard Deviation of 0.0026. The Mean (1.951) multiplied by 1.0%
1.951 x 0.01) = 0.01951, which when added to the 0.005 (0.01951 + 0.005) =
0.02451 OD, which is the Allowed Deviation. Since the Standard Deviation is less
than this value, the reader meets the test criteria.
3. After gathering data for the Linearity Test, the plate is read five more times with the A1
well in the H12 position ("Turnaround" position). This results in values for the four
corner wells that can be used to assess alignment. Calculate results for the Alignment
Test:
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Calculate the means of the wells A1 and H1 in the Normal plate position (data
from Linearity Test) and in the Turnaround position.
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Compare the mean reading for well A1 to its mean reading when in the H12
position. Next, compare the mean values for the H1 well to the same well in the
A12 position. The difference in the values for any two corresponding wells should
be within the Accuracy specification for 96-well plates. If the four corner wells are
within the accuracy range, the reader is in alignment.
Example: If the mean of well A1 in the normal position is 1.902, where the
specified accuracy is ±1.0% ±0.010 OD, then the expected range for the mean of
the same well in the H12 position is 1.873 to 1.931 OD. (1.902 x 1.0% = 0.019 +
0.010 = 0.029, which is added to and subtracted from 1.902 for the range.)
Absorbance Liquid Test 3
Repeatability Specification:
± 1.0% ± 0.005 OD from 0.000 to 2.000 OD
± 3.0% ± 0.005 OD from 2.000 to 2.500 OD
1. The plate is read five times at 340 nm. For each well, calculate the Mean OD and
Standard Deviation of the five readings.
2. For each Mean calculated in step 1, calculate the Allowed Deviation using the
Repeatability specification for a 96-well plate (Mean OD x 0.015 + 0.005). For each well,
its Standard Deviation should be less than its Allowed Deviation.
Synergy LX User Manual
1501000N Revision A BioTek Instruments, Inc.