STAR Protocols (Sep 2023)

A high-throughput protocol for measuring solution pH of bacterial cultures using UV-Vis absorption spectrophotometry

  • Kristen A. Engevik,
  • Hyland Gonzalez,
  • Clay Daniels,
  • Ronald K. Stavert,
  • Numan Oezguen,
  • Melinda A. Engevik,
  • Thomas D. Horvath

Journal volume & issue
Vol. 4, no. 3
p. 102540

Abstract

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Summary: We present a protocol for measuring the pH of cell-free bacterial-conditioned media based on changes in the ultraviolet-visible (UV-Vis) absorbance spectrum using the pH indicator dye litmus. This protocol includes detailed procedures for performing bacterial culturing, examining bacterial growth, collecting cell-free supernatant, litmus dye addition, and pH-based calibration curve preparations. This assay has been designed for flexible formatting that can accommodate both high-volume and low-volume sample sets. : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

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