Biotek Synergy H1, Operator'S Manual

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conducting linear dilutions, an R
2
value of at least 0.9900 is considered
adequate.
2. Calculate the results for Repeatability:
o
Calculate the Mean and Standard Deviation for the five readings taken at each
concentration. Only one row of data needs to be analyzed.
o
For each Mean below 2.000 OD, calculate the Allowed Deviation using the
Repeatability specification for a 96-well plate of ±1.0% ±0.005 OD. (If above
2.000 OD, apply the ±3.0% ±0.005 specification.)
o
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 0.010) = 0.0195, which, when added to the 0.005
(0.0195 + 0.005) = 0.0245 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:
o
Calculate the means of the wells A1 and H1 in the Normal plate position (data
from Linearity Test) and in the Turnaround position.
o
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
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 0.010 + 0.005). For
each well, its Standard Deviation should be less than its Allowed Deviation.
Example: Five readings in well A1 of 0.802, 0.802, 0.799, 0.798, and 0.801 result
in a Mean of 0.8004 and a Standard Deviation of 0.0018. The Mean multiplied by
1.0% (0.8004 * 0.010) equals 0.008, and when added to 0.005 equals 0.013; this is
Synergy H1 Operator's Manual
Absorbance Testing | 81