Kanamycin Selection of Transformants: Selects kanamycin-resistant E. coli carrying KanR plasmids (50 µg/mL kanamycin).

Blue-White Colony Screening: Differentiates recombinant versus empty vectors via lacZ α-complementation using X-Gal (60 µg/mL) and IPTG (0.1 mM) in E. coli.

Insert Verification in Cloning: Identifies successful insertions by white colony phenotype (lacZα disruption) in E. coli lacZΔM15 strains.

Plasmid Maintenance and Propagation: Maintains KanR plasmids during streaking and passaging of E. coli under selection.

Colony Isolation and CFU Enumeration: Plates dilutions to isolate single colonies and count CFUs under kanamycin selection.

Colony Picking for Downstream Workflows: Picks white colonies for colony PCR, miniprep, and sequencing from E. coli screens.

Library Construction and Screening: Screens ligation, Gibson, or Golden Gate libraries in KanR vectors using blue/white selection in E. coli.

Phenotypic Pre-screening of Expression Constructs: Uses IPTG-inducible systems to assess growth/viability phenotypes of lac/T7-lac–regulated KanR constructs on plates in E. coli.

Strain and Plasmid Purity Checks: Re-streaks transformants to verify clonal purity of E. coli carrying KanR plasmids.

Replica Plating for Verification: Confirms selectable markers and insert status by replica plating of E. coli colonies.