SOP for Preparation of Media Containing Sterile Petridishes, Agar Slants and Agar Butts : Pharmaguideline

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SOP for Preparation of Media Containing Sterile Petridishes, Agar Slants and Agar Butts

Standard operating procedure to prepare the sterile media containing petridishes, slants and butts for environmental monitoring and sub-culturing.

1.0 OBJECTIVE

To lay down the procedure for making sterile Petri dishes, agar slants and agar butts.

2.0 SCOPE

This SOP shall be applicable to Quality Control Dept.

3.0 RESPONSIBILITY

Microbiologist

4.0 ACCOUNTABILITY

Sr. Manager Quality Assurance

5.0 PROCEDURE

5.1 Preparation of Petri dishes

5.1.1 Clean and dry the Petri dishes as per the procedure described in SOP for cleaning of glassware used in microbiology.
5.1.2 Sterilize the cleaned and dried Petri dishes by autoclaving at 121ºC for 15 minutes.
5.1.3 Prepare the required quantity of the medium and sterilize by autoclaving at 121ºC for 15 minutes.
5.1.4 Aseptically pour 20 ml of the sterile, molten agar medium, cooled to 40 to 45°C, in each petridish.
5.1.5 Allow the medium to solidify and pre-incubate at the desired temperature for 24 hours before use.
5.1.6 Prepared petridishes, if not used immediately, shall be refrigerated, but shall be used within 15 days of preparation.
Related: SOP for Media Preparation and Growth Promotion Test

5.2 Preparation of agar slants

5.2.1 For the preparation of agar slants, the medium is dispensed in clean and dry test tubes (8 ml in each test tube) and sterilized by autoclaving at 15 lbs pressure (121ºC) for 15 minutes.
5.2.2 After sterilization, the test tubes are kept in a slanting position, in such a way that the slant occupies a little more than half the length of the test tube.
5.2.3 After the medium is solidified, the test tubes are stacked in a vertical position and stored in a refrigerator, until further use.
5.2.4 Frothing and air bubbles should be avoided for preparation of slants. Prepared slants shall be used within 15 days of preparation.

5.3 Preparation of agar butts

5.3.1 For the preparation of agar butts, the medium is dispensed in clean test tubes (5 ml in each test tube) and sterilized by autoclaving at 15 lbs pressure (121ºC) for 15 minutes.
5.3.2 After sterilization, the tubes are kept in a vertical position, until the molten media solidifies.
5.3.3 The tubes are stored in a refrigerator and are to be used within 15 days of preparation.

6.0 ABBREVIATIONS

6.1 SOP: Standard Operating Procedure
6.2 QA: Quality Assurance
6.3 QC: Quality Control
6.4 Dept.: Department
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