Bacterial Viability Assay Protocol

Acetone, Hunan Huihong Reagent Co., Ltd.; Fluorescein diacetate (FDA) and Ortho-nitrophenyl beta-D-galactopyranoside (ONPG) were purchased from Shanghai Yuanye Biotechnology Co., Ltd. Proteinase K, TE buffer, lysozyme, chloroform:isoamyl alcohol (24:1), Tris-saturated phenol:chloroform:isoamyl alcohol (25:24:1), and Acetone were purchased from Beijing Dingguo Biotechnology Co., Ltd. 0.1 mol/L PBS buffer, 10% SDS, 5 mol/L NaCl, CTAB/NaCl, 3 mol/L sodium acetate, and 70% anhydrous ethanol, etc. were configured by the laboratory. FDA stock solution configuration: add 2 mg of FDA to 1 mL of Acetone solution, mix well to obtain 2 mg/mL of FDA stock solution, and store in the dark at −20°C. FDA reaction solution configuration: the FDA stock solution was added to PBS buffer (pH = 7.6) for a final FDA concentration of 10 μg/mL (Li et al., 2023 (link)).