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Measurement of capsule production

FK Fiorella Krapp
AM Andrew R. Morris
EO Egon A. Ozer
AH Alan R. Hauser
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Capsule production was measured by quantification of uronic acid extracted from equivalent volumes of overnight cultures, as previously reported66. Briefly, extracted samples from 500 μL of overnight cultures were resuspended in water and combined with 1.2 mL sodium tetraborate in concentrated sulfuric acid. Samples were boiled for 5 min, followed by the addition of 20 μL 0.15% 3-hydroxydiphenol in 0.5% NaOH, and the absorbance was measured at 520 nm. CPS levels were determined from a standard curve of D-glucuronic acid (Sigma-Aldrich, St. Louis, MO). Samples were normalized to the total viable bacteria in the culture (micrograms uronic acid/106 CFU) and were measured in triplicate.

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