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Surfactant-free Few-Layer-Graphene (FLG) suspensions were prepared by wet media delamination of unmodified isostatic graphite in organic solvents using a shaking plate for fast screening of favorable process parameters and a stirred media mill as a delamination tool.
The cells were washed twice with glucose-free KRH and incubated in this medium ± 20 μM MnTMPyP for 30 min at 37 °C using a shaking plate incubator (Labnet International) set at 100 rpm.
Cells were then incubated in KRH ± 20 μM MnTMPyP (a cell-permeative superoxide dismutase mimetic [21]) for 30 min at 37 °C using a shaking plate incubator (Labnet International, Oakham, UK) set at 100 rpm.
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Fluids were maintained at 37°C using a shaking water bath.
Cola drinks had carbon dioxide content largely removed by vigorous shaking using a stirring plate and placing a magnetic bar in a container filled with the liquid prior to being offered to the animals at room temperature.
Briefly, 10 % BH was mixed with an equal volume of 4%% sarcosyl in PBS, supplemented with 50 mM Tris, pH 7.5, and digested with 20 μg/ml PK (New England BioLabs) for 30 min at 37 °C with 1000 rpm shaking using a DELFIA plate shaker (Wallac) placed in 37 °C incubator.
Absorbance values were recorded after a 5-s shake using a microtitre plate reader at a wavelength of 490 nm.
The plates were covered with a sterile lid, and incubated at 35 °C for 24 h under shaking using a plate shaker (Flow Laboratory Germany) at 300 rpm.
Growth was assessed by Optical Density (OD) at 600 nm until one week incubation at 28°C with shaking (450 rpm) using a spectrofluorometer plate reader (TECAN Infinite M200).
Growth curves were obtained by measuring absorbance at 595 nM in 5-min intervals for 24 hr with shaking at each interval using a BMG Labtech FLUOstar OPTIMA plate reader.
The content of each well was mixed thoroughly with a multi-channel pipette and the macro-well plates were covered with the sterile sealer and incubated at 35 °C for 24 h (for bacteria) and 48 h (for yeasts) under shaking by using a plate shaker (Flow Laboratory, Germany) at 300 rpm.
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