sacB Counterselection: Negative selection for allelic exchange using sacB-based suicide plasmids on Escherichia coli, Pseudomonas aeruginosa, Vibrio cholerae, and Bacillus subtilis.

Protoplast/Spheroplast Osmoprotection: Plating and regeneration of fragile cells and protoplasts (e.g., Bacillus subtilis, Streptomyces spp.) using sucrose for osmotic stabilization without added NaCl.

Salt-Sensitive Bacteriophage Work: Plating and propagation of NaCl-sensitive phages on Escherichia coli lawns using low-ionic LB agar.

Electroporation Recovery: Post-electroporation recovery/plating of delicate Gram-negative cells (e.g., Pseudomonas aeruginosa, Acinetobacter baylyi) requiring osmotic support.

Salt Sensitivity/Halotolerance Testing: Culturing and phenotyping salt-sensitive mutants where NaCl-free medium avoids ionic stress.

Maintenance of sacB-Containing Constructs: Routine culturing and selection steps for strains harboring sacB-based vectors under sucrose pressure.

Osmotic Stress and Envelope Integrity Assays: Evaluating cell envelope mutants under high sucrose, low-ionic conditions.