Growth of DAP-auxotrophic strains: Cultivation and maintenance of E. coli ΔdapA and donor strains (e.g., E. coli BW29427, E. coli WM3064) that require 100 µg/mL diaminopimelic acid.

Bacterial conjugation workflows: Plating donor and mating mixes where the DAP-dependent donor requires supplementation during mobilization of oriT-bearing plasmids (e.g., RP4/RK2 systems).

Donor counterselection by DAP omission: Post-mating selection by replica-plating or streaking onto DAP-free media to eliminate DAP-dependent donors while recovering transconjugants.

Suicide vector delivery and allelic exchange: Conjugative delivery of non-replicative plasmids from DAP-dependent donors for genome editing and allelic replacement in recipients.

Antibiotic selection with donor containment: Combined antibiotic selection on DAP plates to maintain plasmids in DAP-requiring donors while preventing donor overgrowth on DAP-free selective plates.

Colony isolation and CFU enumeration: Streaking and plating to isolate single colonies and quantify viable counts of DAP-dependent strains.

Strain storage and transport: Short-term maintenance and shipment of DAP-auxotrophic donor strains on solid medium without loss of DAP-dependent viability.

Routine cloning workflows with donor strains: Verification, purification, and stabilization of plasmids maintained in DAP-requiring E. coli hosts prior to transfer into recipients.