Biotek Synergy H1, Operator'S Manual

Page: 157 / 190

Test Solutions
Filter solutions to remove particulates that could cause erroneous
readings. Do not allow dust to settle on the surface of the solution; use
microplate covers or seals when not reading the plate.
Wrap the vial containing the MUB stock solution in foil to prevent
exposure to light.
Discard any open, unused solutions after seven days.
1. Prepare the buffer (CBB) solution:
l
(Optional, but recommended) Using a 0.45-micron filter, filter 200 mL of
deionized or distilled water.
l
Open and dissolve the contents of two CBB capsules (do not dissolve the outer
gelatin capsule) into 200 mL of the water.
l
Stir the solution (preferably using a stir table) until the CBB is completely
dissolved.
2. Prepare the MUB stock solution:
l
Add 1 mL of 100% methanol to the 10-mg vial of MUB.
l
Make sure all of the dye has completely dissolved and is well mixed. This yields a
10 mg/mL stock solution.
l
Wrap the solution in aluminum foil to prevent exposure to light.
3. Prepare the dilutions. Label each with “MUB” and the concentration.
Mix this MUB solution: with: to make:
0.5 mL of 10 mg/mL stock solution 4.5 mL of 100% methanol 1 mg/mL
0.88 mL of 1 mg/mL solution 4.12 mL of CBB 176 µg/mL
0.1 mL of 176 µg/mL solution 9.9 mL of CBB 1.76 µg/mL
0.5 mL of 1.76 µg/mL solution 4.5 mL of CBB 176 ng/mL
1 mL of 176 ng/mL solution 9 mL of CBB 17.6 ng/mL
(100 nM)
Synergy H1 Operator's Manual
Fluorescence Liquid Tests | 131