Exact(8)
The engineered C. reinhardtii strains elevated intracellular steady-state ATP levels by more than twofold across their viable temperature range, displayed ~ 3-fold higher growth rate and biomass, and ~ 25% higher lipid/oil accumulation compared to wild-type under both normal and cold growth conditions.
After incubation the cells were washed once with cold growth medium, twice with cold PBS, and resuspended in cold PBS.
The cells were resuspended in a cold growth medium and stained with propidium iodide.
Importantly, prophages in FSL R5-192 do not seem to be interrupting genes related with cold growth.
To assess cold growth, a single colony for each given isolate was first inoculated into BHI broth and grown for 24 h at 32°C.
Different stressful conditions (e.g. acidity, osmolarity, high temperature [ 64, 65]), could induce the prophage lytic cycle, and consequently lyse the cells; experimental evidence will be necessary through to determine whether cold-growth could induce the lytic cycle in the prophage identified here, thus preventing cold growth.
To identify genomic features and genes that may be linked to the unique phenotypic characteristics of clade II strains (e.g. cold growth), we performed an enrichment analysis for RAST subcategories among clade II and III genomes.
For tumor growth experiments on the CAM, 5 × 10 HT29 cells or 7.5 × 10 OE19 cells were resuspended in 50 μL cold growth factor reduced Matrigel (Becton Dickinson, Breda, The Netherlands) and kept on ice until grafting.
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