Biotek Synergy HTX, Operator'S Manual

Page: 136 / 184

114
|
Chapter 6: Instrument Qualification
Alignment Test
1.
Using the plate prepared for the Linearity Test on the previous page,
conduct a Turnaround test by reading the plate
five times
with the A1
well in the H12 position. Save the data after each read.
This test results in values for the four corner wells that can be used to
determine alignment.
2.
Calculate the means of the wells A1 and H1 in the Normal plate position
(data from Linearity Test) and in the Turnaround position (from Step 1
above).
3.
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 the instrument.
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 and subtracted from 1.902 for
the range.)
If the four corner wells are within the accuracy range, the reader is in
alignment.
Absorbance Liquid Test 3 (optional)
Materials
• New 96-well, clear, flat-bottom microplate (Corning Costar #3590 is
recommended)
•
Calibrated hand-pipette(s)
• Beakers and graduated cylinder
• Precision balance with readability to 0.01 g
•
Buffer Solution described below

Read a sample of the buffer solution at 340 nm. This solution should
have an optical density of approximately 0.700 to 1.000. This value is
not critical, but it should be within this range.

If low, adjust up by adding β -NADH powder until the solution is at
least at the lower end of this range. Do not adjust if slightly high.
BioTek Instruments, Inc.