Biotek Synergy HT Operator's Manual page 99

Prepare the Plate
1. Prepare a
with 5 mL of the PBS solution.
2. Prepare a
with 10 mL of the PBS solution.
3. Pipette the three solutions into a
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Read the Plate
1. Using Gen5, read the microplate five times using Normal mode, single
wavelength at 340 nm, no blanking. Save the data after each read.
2. Print out the five sets of raw data, or export them to an Excel spreadsheet.
Analyze the Results
1. For each well, calculate the mean 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 of ±1.0% ± 0.005 OD
(Mean 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
will 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 the
0.005 (0.008 + 0.005) equals
A1. Since the standard deviation for well A1 is less than 0.013, the well
meets the test criteria.
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3. Calculate the results for Linearity:
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Synergy HT Operator's Manual
75% Test Solution
50% Test Solution
150 µL of the 100% Test Solution into all wells of columns 1 and 2
150 µL of the 75% Test Solution into all wells of columns 3 and 4
150 µL of the 50% Test Solution into all wells of column 5 and 6
Repeatability Specification:
± 1.0% ± 0.005 OD from 0.000 to 2.000 OD
± 3.0% ± 0.005 OD from 2.000 OD to 3.000 OD
For each of the three Test Solutions, calculate the mean absorbance for
the wells containing that solution (mean of wells A1 to H2, A3 to H4,
and A5 to H6).
Perform a regression analysis on the data to determine if there is
adequate linearity on a program such as Microsoft
Absorbance Liquid Tests |
by mixing 15 mL of the 100% Test Solution
by mixing 10 mL of the 100% Test Solution
new 96-well microplate:
0.013 , which is the allowed deviation for well
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Excel:
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