how to make nutrient agar
Standard Nutrient Agar No. 1 M1210 Standard Nutrient Agar No. 1 is used for cultivation of fastidious bacteria.
Composition** Ingredients Gms / Litre
Peptone, special 15.000
Yeast extract 3.000
Sodium chloride 6.000
Dextrose 1.000
Agar 12.000
Final pH ( at 25°C) 7.5±0.2 **Formula adjusted, standardized to suit performance parameters Directions Suspend 37 grams in 1000 ml distilled water. Heat to boiling to dissolve the medium completely. Sterilize by autoclaving at 15 lbs pressure (121°C) for 15 minutes. Mix well and pour into sterile Petri plates.
Principle And Interpretation Nutrient media are basic culture media used for culturing maintaining microorganisms
(1), and to check culture purity prior to biochemical or serological testing. Standard Nutrient Agar No. 1 is used for the isolation and enumeration of bacteria. Standard Nutrient Agar No. 1 can be used as a culture media base, when supplemented with blood, ascetic fluids, serum or egg yolk etc which makes it suitable for the cultivation of relatively fastidious organisms
(2). The media can be used for cultivation of Streptococci, Pneumococci and Erysipelothrix species. Quality Control Appearance Cream to yellow homogeneous free flowing powder Gelling Firm, comparable with 1.2% Agar gel. Colour and Clarity of prepared medium Light amber coloured clear to slightly opalescent gel forms in Petri plates Reaction Reaction of 3.7% w/v aqueous solution at 25°C. pH : 7.5±0.2 pH 7.30-7.70 Cultural Response Organism Inoculum (CFU) Growth Recovery Cultural Response Escherichia coli ATCC 25922 50-100 good-luxuriant >=70% Erysipelothrix rhusiopathiae ATCC 19414 50-100 good-luxuriant Listeria monocytogenes ATCC 19111 50-100 good-luxuriant >=70% Staphylococcus aureus ATCC 25923 50-100 good-luxuriant >=70% Streptococcus pneumoniae ATCC 6303 50-100 good-luxuriant >=70%
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Streptococcus pyogenes ATCC 19615 50-100 good-luxuriant >=70% Shigella flexneri ATCC 12022 50-100 good-luxuriant >=70% Storage and Shelf Life Store below 30°C in tightly closed container and the prepared medium at 2-8°C.
Use before expiry date on the label.
Reference 1.Lapage S., Shelton J. and Mitchell J., 1970, Methods in Microbiology, Norris J. and Ribbons D., (Eds.), Vol. 3A, Academic Press, London. 2.MacFaddin J. F., 1985, Media for Isolation-Cultivation-Identification-Maintenance of Medical Bacteria, Vol. I, Williams and Wilkins, Baltimore
