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

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

2023STAR Protocols11 citationsDOIOpen Access PDF

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.

Topics & Concepts

AbsorbanceChemistrySpectrophotometryChromatographyLitmusAbsorption (acoustics)Bacterial cell structureVolume (thermodynamics)Analytical Chemistry (journal)Materials scienceBacteriaBiologyOrganic chemistryPhysicsComposite materialGeneticsQuantum mechanicsAnalytical Chemistry and SensorsBiosensors and Analytical DetectionMicrofluidic and Capillary Electrophoresis Applications
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