As part of this study, we performed an evolution experiment where we selected for colony spreading in eight Bacilli strains and species.. Microbial life cycles link global modularity in regulation to mosaic evolution

We grew eight Bacilli strains and species, four replicates each, on a chemically defined agar growth medium by inoculating cells in the center of the growth plate. These cells were subsequently allowed to form a colony. After a week, cells from the outermost edge of the colony were transferred to a new growth medium, for another growth cycle. In total, cells were grown for 11 consecutive growth cycles. Then, we selected clones from all of the populations at various time points to characterize their phenotypes. For those populations that lost the capacity to sporulate, we sequenced clones both before and after the loss of sporulation in order to identify the causal mutations that resulted in the loss of sporulation.