Biotek Synergy H1, Operator'S Manual

Page: 143 / 190

the plate. When prompted, rotate the plate 180 degrees.
5. When finished:
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Save the experiment. Refer to the instructions on page 80 to perform calculations
and determine pass/fail.
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Troubleshooting tips are provided on page 82 .
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Test descriptions are provided on page 75 .
Absorbance Liquid Test 3
Absorbance Liquid Test 3 is provided for sites requiring proof of linearity at 340 nm. This
test is optional because the Synergy H1 has good "front end" linearity throughout its
wavelength range. As an alternative, the 340 nm Absorbance Test Plate (BTI #7260551)
may be used for this test.
Materials
Manufacturer part numbers are subject to change.
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New 96-well clear flat-bottom microplate (Corning Costar #3590 recommended)
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Calibrated hand pipette(s)
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Beakers and graduated cylinder
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Precision balance with readability to 0.010 g
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Buffer solution described below
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Gen5 protocol Synergy H1 Abs Test 3.prt described on page 79
Buffer Solution
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Deionized water
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Phosphate-Buffered Saline (PBS), pH 7.2–7.6, Sigma tablets, #P4417 (or equivalent)
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β-NADH Powder (β-Nicotinamide Adenine Dinucleotide, Reduced Form) Sigma bulk
catalog number N 8129, or preweighed 10-mg vials, Sigma number N6785-10VL (or
BioTek PN 98233). Store the powder according to the guidelines on its packaging.
1. Prepare a PBS solution from the Sigma tablets.
2. In a beaker, mix 50 mL of the PBS solution with 10 mg of the β-NADH powder and
mix thoroughly. This is the 100% Test Solution .
3. (Optional) Read a 150-μL sample of the solution at 340 nm; it should be within
0.700 to 1.000 OD. If low, adjust up by adding more powder. Do not adjust if
slightly high.
Synergy H1 Operator's Manual
Absorbance Liquid Tests | 117