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The filter papers were frozen in liquid nitrogen for 1 min, and then were placed on top of a pre-soaked filter paper in a 100 mm sterile plate with Z buffer/X-gal solution [2 ml of Z buffer, 6 μl ofβ-mercaptoethanol, 20 μl of X-gal stock solution (40 mg/ml)].
The donor plate was a 96-well filtrate plate (Multiscreen® IP Sterile Plate PDVF membrane, pore size is 0.45 µM, catalog no. MAIPS4510) and the acceptor plate was an indented 96-well plate (Multiscreen®, catalog no. MAMCS9610 from Millipore).
Then, BG-110 medium containing 1.5% agar solution was poured onto a silicon mold that was placed on a sterile plate to produce patterned solid medium with microwells on the bottom (Figure S1B).
The plate was covered with a sterile plate sealer.
A control, consisting in a sterile plate or tube, was also run parallel to each experiment.
200 μL of each mixture was dispensed into each well of a 96-well sterile plate using multi-channel pipette.
Similar(41)
The cells were seeded into 96-well sterile plates at a density of 5 × 103 cells per well and incubated for 24 h overnight.
For the agar dilution method, the 6-mm sterile plates were loaded with 20 μL of test samples of the biosynthesized AgNPs (2.0 mM), AgNO3 (2.0 mM), aqueous longan peel extract, or the penicillin solution (200 μg/mL).
To reduce fungal and bacterial contamination, all experiments were performed using autoclaved medium, sterile plates, pipettes, microcentrifuge tubes and ethanol-rinsed utensils.
The HTS assay was carried out in 1536-well sterile plates (Greiner, 782074).
Cells were seeded and adhered onto appropriate sterile plates for 24 h before treatments.
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