Nitrate Reduction Essay

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Sigma-Aldrich Chemie GmbH · Industriestrasse 25 · Postfach · CH-9471 Buchs / Switzerland Tel. +41 / 81 755 25 11 · Fax +41 / 81 756 54 49 · flukatec@sial.com 73426 Nitrate Reduction Test Bacterial species may be differentiated on the basis of their ability to reduce nitrate to nitrite or nitrogenous gases. The reduction of nitrate may be coupled to anaerobic respiration in some species. Kit content 1. Nitrate Broth (Fluka 72548) Peptone Meat extract Potassium nitrate 5 g/l 3 g/l 1 g/l Qty. 100 g Final pH 7.0 ± 0.2 at 25°C 2. Sulfanilic acid (Fluka 86090) = Reagent A 3. N,N-Dimethyl-1-naphthylamine (Sigma D4011) = Reagent B 4. Zinc (Fluka 93027) (all kit components are available as single components) 100 g 10 ml 1g Preparation of Reagents: Sulfanilic acid solution (Reagent A): Dissolve 8 g of sulfanilic acid in 1 litre 5N acetic acid. Store Reagent A at room temperature for up to 3 months, in dark. Reagents may be stored in dark brown glass containers; bottles may be wrapped in aluminum foil to ensure darkness. α-Naphthylamine solution (Reagent B): Dissolve 6 g of N,N-Dimethyl-1-naphthylamine in 1 litre 5N acetic acid. Store Reagent B at 2 to 8°C for up to 3 months, in dark. Reagents may be stored in dark brown glass containers; bottles may be wrapped in aluminum foil to ensure darkness. Directions: Dissolve 9 g of Nitrate Broth in 1 litre distilled water. Dispense 10 ml aliquots of the broth into tubes fitted with Durham tubes. Sterilize by autoclaving at 121°C for 15 minutes. Inoculate the tubes heavily with a fresh culture of the suspect organism. Inoculate at least 1 ml sample in a tube or take a big part of a colony with an inoculating loop. Do not forget a negative control without any bacteria. A successful nitrate reduction test is dependent on performing the test under the correct conditions. That means the organisms needs the accurate growth media, the

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