TRI Reagent® Sigma

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	Helicobacter pylori strains (Table ​Table11) were grown with gentle agitation (120 rpm) in 30 ml of Brucella broth at 37°C until mid-exponential phase (OD = 0.7). For heat-shock treatment, the wild type (WT) culture was split into 15 ml-aliquots and one sample was subjected to heat-shock at 42°C for 30 min (heat-shock sample, HS). A volume of 10 ml cell culture was then added to 1.25 ml of ice-cold EtOH-phenol stop solution (5% acid phenol, in EtOH) to stop growth and prevent RNA degradation. Cells were pelleted, stored at −80°C, and then used to extract total RNA with TRI-reagent (Sigma-Aldrich), according to manufacturer’s protocol.

Protocol tips

Helicobacter pylori strains (Table ​Table11) were grown with gentle agitation (120 rpm) in 30 ml of Brucella broth at 37°C until mid-exponential phase (OD = 0.7). For heat-shock treatment, the wild type (WT) culture was split into 15 ml-aliquots and one sample was subjected to heat-shock at 42°C for 30 min (heat-shock sample, HS). A volume of 10 ml cell culture was then added to 1.25 ml of ice-cold EtOH-phenol stop solution (5% acid phenol, in EtOH) to stop growth and prevent RNA degradation. Cells were pelleted, stored at −80°C, and then used to extract total RNA with TRI-reagent (Sigma-Aldrich), according to manufacturer’s protocol.

Publication protocol

Helicobacter pylori strains (Table ​Table11) were grown with gentle agitation (120 rpm) in 30 ml of Brucella broth at 37°C until mid-exponential phase (OD = 0.7). For heat-shock treatment, the wild type (WT) culture was split into 15 ml-aliquots and one sample was subjected to heat-shock at 42°C for 30 min (heat-shock sample, HS). A volume of 10 ml cell culture was then added to 1.25 ml of ice-cold EtOH-phenol stop solution (5% acid phenol, in EtOH) to stop growth and prevent RNA degradation. Cells were pelleted, stored at −80°C, and then used to extract total RNA with TRI-reagent (Sigma-Aldrich), according to manufacturer’s protocol.

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