Bacteriological Analytical Manual
|*Potassium cyanide (KCN)||0.5 g|
|Proteose peptone No. 3 or polypeptone||3 g|
|Distilled water||1 liter|
Dissolve above ingredients except potassium cyanide and autoclave 15 min at 121°C. Cool and refrigerate at 5-8°C. Final pH, 7.6 ± 0.2.
Prepare KCN stock solution by dissolving 0.5 g KCN in 100 ml sterile distilled water cooled to 5-8°C.
Using bulb pipetter, add 15 ml cold KCN stock solution to 1 liter cold, sterile base.
DO NOT PIPET BY MOUTH!!!
Handle with gloves!!
Mix and aseptically dispense 1.0-1.5 ml portions to 13 × 100 mm sterile tubes. Using aseptic technique, stopper tubes with No. 2 corks impregnated with paraffin. Prepare corks by boiling in paraffin about 5 min. Place corks in tubes so that paraffin does not flow into broth but forms a seal between rim of tubes and cork. Store tubes at 5-8°C no longer than 2 weeks before use.
Retain no longer than two weeks before use.
Original Source: Bacteriological Analytical Manual, 8th Edition, Revision A, 1998.