February 2013 Text revised, formulation is unchanged from previous (March 2006 version)
|Trypticase soy broth||30 g|
|Yeast extract||6 g|
|Monopotassium phosphate (anhydrous)||1.35 g/liter|
|Disodium phosphate (anhydrous)||9.6 g/liter|
|Sodium Pyruvate (Sodium salt)||1.11 g/liter|
|Distilled water||1 liter|
Weigh ingredients and dissolve in water. Autoclave 15 min at 121°C. Final pH, 7.3 ± 0.1.
[Note: Optionally a filter-sterilized 10% (w/v) sodium pyruvate solution may be added after autoclaving (11.1ml/L)]
|Acriflavin HCl||10 mg/liter|
|Nalidixic acid (sodium salt)||40 mg/liter|
Prepare acriflavin and nalidixic acid supplements as 0.5% (w/v) stock solutions in distilled water. Prepare cycloheximide supplement as 1.0% (w/v) stock solution in 40% (v/v) solution of ethanol in water. Filter-sterilize stock solutions and store at 4°C, protect acriflavin from light.
Aseptically add the 3 selective supplements to enrichment after 4 h incubation at 30°C as described in Chapter 10. Final volume per supplement stock solution determined by total volume of BLEB used in enriched sample.
Original Source: Bacteriological Analytical Manual, 8th Edition, Revision A, 1998.