Preparation and Disposal of Nutrient Agar Plates

Introduction This method may be used to prepare nutrient agar plates for short term usage where meticulous sterility is not required. If plates are to be kept for any length of time before use, then the agar solution should be sterilised at 120°C in a pressure cooker. The procedure for safe disposal of used agar plates is also described Procedure for Preparation 1. Weigh out 12.6g nutrient agar. 2. With gentle stirring on a hotplate-stirrer, slowly add the agar to 450ml of cold demineralised water in an 800ml beaker. 3. When addition of the agar is complete, heat the mixture with vigorous stirring and boil for 15 minutes. 4. Remove the beaker from the stirrer hotplate, retrieve the stirrer “flea” using a magnet, and cover the beaker (foil). 5. Allow to cool to 50°C. 6. Pour the agar into clean Petri dishes. 7. Store the plates in the refrigerator (short term only). Hint: Pour the agar to cover about ⅔ of the plate and then gently tip the Petri dish to give a complete covering. In this way there should be sufficient agar for a class set of 30 plates. Procedure for Disposal
(using a Tower brand domestic pressure cooker)

1. Set up the equipment as shown in figure 1. Note: the “biohazard” bag is trimmed down so that it does not interfere with the pressure relief mechanism in the lid of the pressure cooker. The water is outside the bag. 2. Fit the lid of the pressure cooker. 3. Heat strongly on a gas ring until the correct pressure is achieved – the indicator valve will show red. This may occur when most of the water has boiled off and may take twenty minutes or more from moment that steam starts escaping from the relief valve. The gas can then be turned down to the minimum required to keep the indicator showing red. 4. Continue heating to maintain the pressure for a further 15 minutes. 5. When cool, the plates may be disposed of as general waste.
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2007 Page 2 of 2 /var/www/apps/conversion/current/tmp/scratch16913/88971869.doc .Figure 1 indicator valve relief valve lid “biohazard” bag pressure cooker used plates 450ml water small trivet P.M.12. 3.

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