Yeast Two-Hybrid (Y2H) Selection: Double-dropout (-Leu, -Trp) base to select and maintain bait (TRP1) and prey (LEU2) plasmids before reporter screening.

Plasmid Maintenance in Saccharomyces cerevisiae: Selects for LEU2 and TRP1 auxotrophic markers in defined media with 2.0% glucose and phosphate buffering.

Transformant Screening: Rapid selection of S. cerevisiae colonies carrying LEU2/TRP1-marked vectors after transformation.

Library Construction and Pre-Enrichment: Maintains combinatorial plasmid libraries prior to higher-stringency reporter selection (e.g., -His, -Ade).

CRISPR/Cas9 Workflows: Selection of donor or Cas9/sgRNA plasmids using LEU2/TRP1 markers in defined SD-CAA medium.

Mating and Diploid Selection: Retention of complementary LEU2/TRP1 plasmids in mated diploids for downstream interaction assays.

Growth and Phenotyping Assays: Controlled growth of yeast in defined conditions (2.0% glucose; ammonium sulfate nitrogen source) for comparative studies.

Spotting and Serial Dilution Tests: Quantitative fitness and sensitivity assays on -Leu/-Trp dropout backgrounds (with agar added for plates).

Protein–Protein Interaction Mapping: Pre-selection medium for validating plasmid presence before reporter-based interaction readouts.

Plasmid Stability Studies: Monitoring retention/loss of LEU2/TRP1 vectors under selective pressure in defined media.