ASKA Collection Evolution Experiment

Starting Lines

1. Revive each strain by inoculating a scrape of frozen culture from a well of a microplate into 3 ml of 0.1x LB + 30 μg/ml chloramphenicol. Grow overnight.
2. Precondition to the growth medium by transferring 11.7 μl of each culture into 3 ml of fresh 0.1x LB + 30 μg/ml chloramphenicol. Grow overnight.
3. Found evolutionary independent replicates of each strain by initiating one culture for each experimental replicate from a small number (100-1000) of cells. If you are founding three strain mixtures, you need three copies of each strain that will be present in each mixture at this step. Grow for 2 days to ensure that each strain reaches a similar saturating density from the small inoculum.
4. Now begin the evolution experiment. Mix together equal volumes of each culture. Transfer from this mixture the daily dilution into fresh media and grow that culture overnight.