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Spot plating assay

JF Jan Felix
CS Claire Siebert
JD Julia Novion Ducassou
JN Jérôme Nigou
PG Pierre Simon Garcia
AF Angélique Fraudeau
KH Karine Huard
CM Caroline Mas
CB Céline Brochier-Armanet
YC Yohann Couté
IG Irina Gutsche
PR Patricia Renesto
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To accurately compare the oxidative stress responses of the different strains, two-fold serial dilutions of the drugs were prepared in MMH using 2 mL Eppendorf tubes, then dispensed in a 96 well plate (100 µL/well). Exponential phase bacteria diluted to a OD600nm of 0.2 were then added (100 µL) in the wells containing drug dilutions. Posititive (without drug) and negative (without bacteria) controls were also included. Following 1 h incubation at 37 °C in a 5% CO2 incubator without shaking, 3 μL aliquots were spotted PVX-CHA plates further incubated at 37 °C for 48 h to 72 h before being photographed. All strains were processed in parallel in each experiment which included two independent replicates spotted twice.

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