BacTiter-Glo™ Microbial Cell Viability Assay

Protocol tips

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	Add a volume of BacTiter-Glo™ Reagent equal to the volume of cell culture medium present in each well. Mix contents briefly on an orbital shaker and incubate for five minutes.

Protocol tips
Add a volume of BacTiter-Glo™ Reagent equal to the volume of cell culture medium present in each well. Mix contents briefly on an orbital shaker and incubate for five minutes.

Publication protocol

S. epidermidis BCRC-11030 (ATCC 12228) was obtained from Bioresource Collection and Research Center (Hsinchu, Taiwan) and was cultured on Nutrient agar (Oxoid, Hampshire, England). Single colonies were inoculated in Nutrient broth and cultured at 37°C until reaching around OD600 1.0 (logarithmic growth phase) under shaking conditions. Then the seed culture was diluted with fresh Nutrient broth to the value of 0.1 (optical density (OD)600) for further microbial growth experiments. Drugs and drug-contained microemulsions were diluted by dimethyl sulfoxide (DMSO) in a 2-fold serial manner and added at the 1% volume ratio to the 96-well microtiter plates. The bacteria were cultured in plates (volume=0.2mL) for 24h and the growth was quantified by the BacTiter-Glo™ assay (Promega). The assay uses bioluminescence produced by luciferase to detect the intracellular concentration of ATP, which correlates with viable bacterial cell numbers. 16) In brief, the reaction involves adding a single reagent directly to bacterial cells cultured in medium and measuring luminescence generated by bacterial ATP and a thermostable luciferase. The bioluminescence was measured by Epoch Spectrophotometer (Biotek, Winooski, VT, U.S.A.). The control group was S. epidermidis incubated with 1% (v/v) DMSO.

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Manufacturer protocol

Download the product protocol from Promega for BacTiter-Glo™ Microbial Cell Viability Assay below.

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