Arachidonic Acid Growth Inhibition Assay

Overnight cultures of JE2, tcaA, and lcpA mutants were pelleted, and the supernatant saved. Growth inhibition assays were subsequently performed by adding 20 μl of OD_600nm 0.1 bacterial suspension to 180 μl 10% overnight supernatant diluted in fresh MHB++ containing 400μM of arachidonic acid (Sigma). Purified WTA extracts were also added to the growth media at a final concentration of 2%. The ability of the bacteria to survive the arachidonic acid was determined by quantifying bacterial growth (OD_600nm) following 24 h at 37°C using a CLARIOstar plate reader (BMG Labtech).

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Douglas E.J., Palk N., Brignoli T., Altwiley D., Boura M., Laabei M., Recker M., Cheung G.Y., Liu R., Hsieh R.C., Otto M., O’Brien E., McLoughlin R.M, & Massey R.C. (2023). Extensive re-modelling of the cell wall during the development of Staphylococcus aureus bacteraemia. bioRxiv.

Publication Preprint 2023

arachidonic acid CLARIOstar plate reader

Arachidonic acid Assays Bacterial Growth media Inhibition

Corresponding Organization : University College Cork

Other organizations : University of Bath, University of Milan, University of Exeter, University of Tübingen, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Trinity College Dublin

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Variable analysis

independent variables

Bacterial strains (JE2, tcaA, and lcpA mutants)

dependent variables

Bacterial growth (OD600nm)

control variables

Overnight supernatant dilution (10%)
Arachidonic acid concentration (400 μM)
Purified WTA extract concentration (2%)
Incubation time (24 h)
Incubation temperature (37°C)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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