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

STAR Protoc. 2023 Sep 15;4(3):102540. doi: 10.1016/j.xpro.2023.102540. Epub 2023 Sep 7.

Abstract

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.

Keywords: High Throughput Screening; Microbiology.

MeSH terms

  • Calibration
  • Hydrogen-Ion Concentration
  • Light*
  • Spectrophotometry / methods
  • Spectrophotometry, Ultraviolet / methods