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Antimycobacterial activity screening by Resazurin Assay: M. tuberculosis H37Rv was grown in Middlebrook 7H9 broth (Difco BBL, Sparks, MD, USA) supplemented with 10% OADC (Becton Dickinson, Sparks, MD, USA). The culture was diluted to McFarland 1 standard with the same medium. From this, 50 μl of this culture was added to 450 μl of fresh medium in 2 ml microcentrifuge tubes. Stock solutions (2 mg/ml) of the test compounds were prepared in dimethylformamide (DMF). The compounds were tested at 1, 10 and 50 μg/ml concentrations. Control tubes had the same volumes of DMF without any substrate. After incubation at 37 °C for 7 days, 20 μl of 0.01% Resazurin (Sigma, St. Louis. MO, USA) in water was added to each tube. Resazurin, a redox dye, is blue in the oxidized state and turns pink when reduced by the growth of viable cells. The control tubes showed a colour change from blue to pink after 24 h at 37 °C. Compounds which prevented the change of colour of the dye were considered to be inhibitory to M. tuberculosis.
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