Chocolate agar media, is a non-selective, enriched growth medium used for the isolation of pathogenic bacteria like Haemophilus influenza or Neisseria spp. The name of the medium is derived from its chocolate-brown colored appearance which is obtained by the lysis of red blood cells. The preparation of chocolate agar is done by slowly heating the blood agar medium containing red blood cells to a temperature of 80oC for initiating RBC lysis. The lysis of red blood cells allows release of growth factors like nicotinamide adenine dinucleotide (factor V or NAD) and hemin (factor X) into the medium which are utilized by the fastidious bacteria for their luxuriant growth. The medium is recommended for isolating microorganisms from clinical specimens like blood, vaginal or endocervical samples. With addition of supplements, Chocolate agar base acts a selective medium that supports the excellent growth of gonococcus when isolated from a mixed flora.