Primary Isolation of Mycobacteria: Selective cultivation of slow-growing species such as Mycobacterium tuberculosis complex from clinical specimens using Middlebrook 7H10 base with 0.5% glycerol and 10% OADC enrichment.

Subculture from Liquid Media: Plating of positive 7H9/MGIT cultures to obtain isolated colonies of Mycobacterium spp. for downstream testing.

Colony Morphology and Pigmentation: Assessment of colony texture, rough/smooth variants, and chromogenicity for differentiating Mycobacterium species.

Enumeration (CFU Counting): Quantitative plating to determine viable counts of Mycobacterium cultures for growth kinetics and standardization.

Antimicrobial Susceptibility Testing (Agar Proportion): Preparation of drug-containing plates and controls to evaluate susceptibility of M. tuberculosis complex and NTM.

Recovery from Decontaminated Specimens: Growth of mycobacteria post-NaOH/NALC processing aided by OADC enrichment and malachite green selectivity.

Species Differentiation and Identification Workflows: Isolation for downstream PCR/NGS, biochemical panels, and MALDI-TOF from pure Mycobacterium colonies.

Maintenance and Stock Culture: Short- to medium-term storage and passaging of reference strains (e.g., M. tuberculosis, M. bovis BCG, M. avium) on a nutrient-rich solid matrix.

Environmental and NTM Isolation: Culturing non-tuberculous mycobacteria such as M. avium complex and M. kansasii from water, soil, and biofilm samples.

Contamination Screening: Detection of non-mycobacterial contaminants during mycobacterial culture workflows utilizing malachite green inhibition.