I am attempting to screen for phosphate solubilizing bacteria, using a modified pikovskaya's broth as a growth medium. After incubation/shaking for 2 days, the cultures are centrifuged at 12,000 rpm for 15 minutes to remove any insoluble phosphorus and/or bacterial cells from the supernatant. This supernatant is then sampled for reaction with a molybdate based coloring reagent. When carrying out the molybdate-blue method, my uninoculated broth appears to have as much coloration as my expected PSB containing broth. I know that the reagents are not at fault because they produce a linear standard curve with monopotassium phosphate dilutions. Why would my broth which should contain no soluble phosphate change colors?

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