Chocolate agar is an enriched culture medium specifically formulated for the isolation of fastidious organisms, particularly Neisseria and Haemophilus species. The medium is prepared by heating 5-7% horse blood in GC agar base to release X factor (hemin) from hemoglobin, essential for Haemophilus growth. The addition of IsoVitaleX enrichment provides V factor (NAD) along with a defined mixture of growth-enhancing components including vitamins, amino acids, coenzymes, dextrose, and ferric ions, which collectively support the optimal growth of pathogenic Neisseria species.