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Serial dilutions of the bacteria were plated onto nutrient agar in triplicate and incubated at 37 °C for 18 h.
The bacteria were plated on Müeller Hinton Agar by the spread-plate technique to obtain single colony.
The bacteria were plated on imMedia Zeo Agar plates (Invitrogen).
For growth on solid medium, bacteria were plated on MB7H11(Difco, BD) agar supplemented with OADC.
Cells were lysed and bacteria were plated as described above for invasion and adherence assays.
The conjugative bacteria were plated onto BCYE agar and incubated at 32°C overnight.
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In this assay, internalized bacteria are plated and counted 24 hours after host cell lysis.
The expression of NfsB in this system sensitises the bacteria to CB1954, resulting in the inhibition of colony formation when the bacteria are plated on agar containing CB1954.
In addition the plant cells were lysed by incubation for 10 min at room temperature in lysis buffer (0.2% Triton X-100, 100 mM potassium phosphate buffer, pH 7.8), and the attached bacteria were plate-counted as described [49].
For isolation of other bacteria, samples were plated on Columbia agar (Oxoid, Basingstoke, UK) with 5% sheep blood and incubated at 37°C in 5% CO2.
In addition, 100 µL of undiluted mineral-bacteria suspensions were plated directly on LB agar.
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