Gbox chemi xx6 gel imaging system

Intact yeast chromosomal DNA was prepared as previously described [101 (link)]. Briefly, cells were grown overnight, and a volume equivalent to an OD₆₀₀ of 6 was washed in 50 mM EDTA and resuspended in 20 μl of 10 mg/ml Zymolyase 100T (Amsbio #120493–1) and 300 μl of 1% Low Melt agarose (Biorad # 1613112) in 100 mM EDTA. Chromosomes were separated on a 1% Megabase agarose gel (Bio-Rad) in 0.5X TBE using a CHEF DRII apparatus. Run conditions were as follows: 60-120s switch at 6 V/cm for 12 hours followed by a 120-300s switch at 4.5 V/cm for 26 hours at 14°C. The gel was stained in 0.5x TBE with ethidium bromide (0.5 μg/ml) for 60 minutes and destained in water for 30 minutes. Chromosomes were visualised using a Syngene GBox Chemi XX6 gel imaging system.

Overnight liquid cultures were diluted to an OD₆₀₀ of 4, serially diluted 1:5 and spotted into agar plates with and without indicated additives using a replica plater (Sigma Aldrich, #R2383). Images of the plates were taken using Syngene GBox Chemi XX6 Gel imaging system. Experiments were performed in 3 biological replicates.