LBL04002

Using the Autoclave for Media Preparation

Author: Tolland	Approved by: S. Thompson	SOP No. LBL04002
Signed:	Signed:	Effective from:
Date:	Date:	Last edited:

1. Purpose This describes the operation of the Melag EuroKlav 29V-S. The autoclave is used for two main procedures:

   i) sterilization of media, glassware and autoclavable equipment prior to another protocol
  ii) Deactivation of GMOs, and GMO exposed waste and equipment.

2. Scope

This should be observed every time the autoclave is used within the LBL BioLab.

3. Responsibilities

The operator of the autoclave is responsible for their own safety and that of others in the vicinity during its use. The particular risks presented in using this piece of equipment are primarily due to the high temperatures generated, steam and high pressures.

The operator is responsible for ensuring complete sterilization of containers, glassware, tubes, media and equipment. To ensure environmental microbes do not contaminate the target microbe strain.

4. Materials

The main consumables for the autoclave are RO water and autoclave tape.

RO water needs to be regularly topped up, into the hatch on the top of the machine.

The autoclave tape should be used to seal items to be sterilized and is stored on the shelf nearby. Order replacement from Sigma or appropriate similar product.

5. Related documents

http://www.phytotechlab.com/pdf/SterilizingNutrientMedia.pdf

6. Definitions

None given

7. Procedures

7.1 Sterilization of liquid media

7.1.1 For sterilization of liquid media up to 1 litre, that requires 15 minutes @ 15 psi and 121 C, please use the "Gentle Programme";

7.1.2 For sterilization of liquid media of greater volumes than 1 litre, you need to preheat the autoclave and place the material to be sterilized in the autoclave 15 minutes before starting the programme. This is to ensure that the bulk volume of liquid comes up to temperature in time for the main sterilization phase.

7.1.3 Then run for 15 minutes @ 15 psi and 121 C using the "Gentle Programme";

7.2 Deactivation of liquid and solid GMO waste ==

7.2.1 Transport the waste to the autoclave in a non-drip autoclavable tray 7.2.2 Ensure there are no folds or containers which can trap air 7.2.3 Sterilize the load using the Universal-Programme (2.1 bar/134 C/15 mins) 7.2.4 once the programme has run, the waste should be transferred to the chemical kill bin, for final deactivation

8. Resources

- Operating manual: http://www.eickemeyer.nl/images/products/2281/500032_Melag_Euroklav_23VS.pdf