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2 mM riboflavin (0.1 mL) was added in 3 mL of reaction mixture (13.33 mM methionine, 75 M NBT, 0.1 mM EDTA, 50 mM phosphate buffer (pH 7.8), 50 mM sodium carbonate, 0.1 mL enzyme extract) and placing the tubes under two 15 W fluorescent lamps for 15 min to start the reaction