1.0 OBJECTIVE:
To lay down a standard procedure for preparation and sterilization of microbiological culture media.
2.0 SCOPE:
This Standard Operating Procedure is applicable for media preparation and sterilization in Microbiology laboratory of Quality Control Department.
3.0 RESPONSIBILITY:
Microbiologist shall be responsible to follow the procedure mention in this SOP.
4.0 ACCOUNTABILITY:
Department Head & QA Head shall be accountable for compliance of this SOP.
5.0 ATTACHMENTS:
Media preparation record –Attachment-I
6.0 PROCEDURE:
6.1 General precautions:
6.1.1 Take clean, dried glass conical flask/borosilicate glass bottle of desired volume as per quantity of
media.
6.1.2 Use a separate spatula for separate media to avoid cross contamination.
6.1.3 Clean the balance in between successive weighing.
6.2 Preparation and sterilization:
6.2.1 Use dehydrated culture media from Merk, Hi Media Laboratories Ltd or any of the Authentic Source.
Check the in house use before date prior to use of the media.
6.2.2 Take clean dried conical flask/borosilicate glass bottle of desired volume as per quantity of media.
Transfer the half of the volume of purified water of required quantity in the conical flask.
6.2.3 Weigh required quantity of de-hydrated media as per volume required, transfer in conical flask and
re-hydrate it as per manufacturer instructions. Make up the volume with remaining quantity of water.
6.2.4 Mix well and heat gently to dissolve the media. Note down the weight taken for each medium.
6.2.5 Check the pH of the media before sterilization. Adjust the required pH (specified on the respective
medium pack) by using either 0.1 M HCl or 0.1 M NaOH solution if required.
6.2.6 In case of broth medium, distribute the suitable quantity of the broth medium in appropriate glass
bottle or test tubes.
6.2.7 Plug the bottles with screw cap or test tubes with a cotton plug or screw cap, wrap with aluminum
foil.
6.2.8 Sterilize the media by autoclaving at temperature121°C and 15 psi/kg/cm² pressure for15 minutes or
as per manufacturer instructions.
6.2.9 After sterilization, use the media for desired purpose and Store the sterilized broth media below 25°C
till further use.
6.2.10 For agar media, after auto claving, cool the medium to about 45-50°C then pour the media
aseptically into sterile Petri plates in 15 to 20 ml quantity in each plate. Allow the plates to solidify.
6.2.11 Close the plates invert them and incubate them for desired purpose.
6.2.12 After preparation of media, record the following details in the format of media preparation record in
format no. SOP/ML/005/F01-00.
6.2.13 Name of media, Batch No., Use before, Date of preparation, Total weight taken, Total volume, pH
before sterilization and Prepared by.
6.2.14 All the media prepared in a day should be assigned a respective batch no. and these batch nos.
should be entered in the media preparation, utility & destruction record.
6.3 Assigning batch/lot number:
6.3.1 All the media prepared in one autoclaved cycle should have respective batch numbers and these
batch numbers should be entered in media preparation record.
6.3.2 Batch no. of autoclaved media should be allotted in the following manner:
XXX/ZZZZZZZ
Where XXX : Autoclave load no. starting from 001 for every year
And ZZZZZZZ: Batch no. of media provided by the manufacturer.
6.4 Storage of rehydrated sterilized media:
6.4.1 Broth medium : Stored below 25°C
6.4.2 Molten agar media : Stored between 55-60°C
6.4.3 Prepared agar plates : Stored below 25°C
7.0 REFERENCES:
In-house
8.0 ABBREVIATIONS:
SOP : Standard operating procedure
°C : Degree Celsius
Psi : Pound per square inch
Ml : Milliliter
HCl : Hydrochloric acid
NaOH : Sodium Hydroxide
CC No. : Change Control number
QA : Quality Assurance
QC : Quality Control
9.0 DISTRIBUTION LIST:
Quality Assurance Department
Quality Control Department
10.0 HISTORY OF REVISION:
| Version Number | Effective Date | Reason for Revision |
| 00 | 20/04/17 | New SOP |
| Title | : | Media preparation record | |||
| Version No. | : | 00 | Page No. | : | 1 of 1
|
| Effective Date | : | Review Date | : | ||
Attachment-I
| S. No. | Date | Name of Media | Media Quantity taken | Volume prepared (ml) | pH before sterilization | Autoclave lot. no./Media batch/lot no. | Prepared by | Checked by |
