Range expansion experiments of one-way competing consortia on solid agar
This protocol is extracted from research article:
Spatial interference scale as a determinant of microbial range expansion
Sci Adv, Nov 21, 2018; DOI: 10.1126/sciadv.aau0695

SRI+, SRI−, LRI+, and LRI− strains were separately grown overnight (16 hours) in LB/Cm medium to stationary phase. Then, cells were subinoculated to 5 ml of LB/Cm medium supplemented with 1× CopyControl Induction Solution in a 50-ml Falcon tube at 1:10 dilution and incubated at 37°C with shaking at 225 rpm for 5 hours. Toxin-producing and sensitive cells were adjusted to an OD600 of 1.0 using fresh medium and mixed at 1:1 ratio for different combinations. Aliquots (1 μl) of each combination were spotted onto LB/Cm agar (15 ml of 1.5% agar in a 90-mm plate) containing arabinose [0.2% (w/v)] and incubated at 37°C. Green and red fluorescence of the colonies were imaged using a Zeiss Axio Zoom.V16 microscope with an AxioCam HRm camera at 12, 24, 36, 48, 72, and 96 hours. In addition, toxin-producing and sensitive cells were mixed at total OD600 = 1 but with different initial ratios (5:1 to 1:5) and with 1:1 ratio but at different initial total OD600. The cell mixtures were spotted onto an agar plate as above and incubated at 37°C for imaging at 80 hours.

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