MEDIA PREPERATION

PROCEDURE

• INTRODUCTION

Ø            Before employing any dehydrated media for analysis ensure that the growth promotion test has been performed for that particular batch or lot or any new media procured.

Ø            Media for Microbiological testing should be prepared as per requirements.

Ø            For preparing any culture media (Solid or Liquid) take properly cleaned glass container of different volumes as per requirement.

Ø            Select the glass container such that the medium volume be prepared, is half of its capacity.

Ø            Follow the instructions given by the manufacturer for   preparing the culture media (Name of the Mfr.Hi-media Laboratory Pvt. Ltd.).

Ø            Weigh the media powder as mentioned on the label of the container and Annexure. Reconstitute with distilled water and mix the powder stirring with glass rod.

Ø            Warm the powder mixed distilled water, if required, to dissolve the powder completely & check the pH of the solution with the help of pH indicator strip.

Ø            Plug the mouth of the container by non-absorbent cotton and wrap butter paper /Alue. Foil to prevent any Contamination and label.

Ø            Boil or sterilize the media at 121°C for 20 min, as per manufacturer instruction and check the pH after sterilization.

Ø            After completion of  all the above procedures label each media containers with media name, media lot no.  date of preparation, and Use before date on the container .

Ø            Pre-incubation must be done at 30-35°C for at least 48 hrs before taken for use. In case if the medium is required to be use immediately, keep appropriate negative and positive control to ensure the proper sterilization and fertility of the medium.

Ø            Record the media preparation and pre-incubation details in Media preparation record as mentioned in Annexure

Ø            Prepared media can be used within 1month period if properly sealed and stored at temperature < 25°C.

Ø            Whenever new bottle is opened write the date of opening, Date of growth promotion test and signature  on the bottle.

List of Media

S.no.	Media	Gms. per Liter	pH	Procedure for preparation
1.	Soyabean Caesin Digest Media (SCDM),	30.00	7.3 ± 0.2	Sterilization at 1210C for 20 min
2.	Fluid Thioglycollate Media (FTG   M),	29.75	7.1 ± 0.2	Sterilization at 1210C for 20 min
3.	Soyabean Caesin Digest Agar     (SCDA)	40.00	7.3 ± 0.2	Sterilization at 1210C for 20 min
4.	Brilliant Green Agar (BGA),	58.00	6.9 ± 0.2	Sterilization at 1210C for 20 min
5.	Bismuth Sulphite Agar(BSA)	52.33	7.7 ± 0.2	Boiling
6.	Deoxycollate Citrate Agar (DCA)	70.52	7.5 ± 0.2	Boiling
7.	Selinite  Broth (SCB),	23.00	7 .0 ± 0.2	Boiling
8.	Tetra thionate Broth (TTB),	77.4 / 980ml	--	Boiling
9.	Mac Conkey Broth Purple (MB),	35.0	7 .3 ± 0.2	Sterilization at 1210C for 20 min
10.	Xylose Lysine Deoxycholate Agar (XLDA),	56.68	7.4 ± 0.2	Sterilization at 1210C for 20 min
11.	Mac ConkeyAgar (MA),	48.5	7.1 ± 0.2	Sterilization at 1210C for 20 min
12.	Sabaurud Dexterose Agar (SDA),	65.0	5.6 ± 0.2	Sterilization at 1210C for 20 min
13.	Azide Dextrose Broth (AZB),	34.7	7.2 ± 0.2	Sterilization at 1210C for 20 min
14.	Vogel Jhonson Agar (VJA)	61.0	7.2 ± 0.2	Sterilization at 1210C for 20 min
15.	Urea Broth (UB),	18.7 / 950ml  DW	6.8 ± 0.2	Sterilization at 1210C for 20 min
16.	Manitol Salt Agar (MSA),	111.0	7.4 ± 0.2	Sterilization at 1210C for 20 min
17.	Baired Parker Agar (BPA),	63.0	7.0 ± 0.2	Sterilization at 1210C for 20 min
19.	Nutrient Broth (NB),	13.0	7.4 ± 0.2	Sterilization at 1210C for 20 min
20.	Cetrimide Agar (CA)	45.3	7.2 ± 0.2	Sterilization at 1210C for 20 min
21.	Antibiotic assay medium no.-3	17.5	7.0 ± 0.2	Sterilization at 1210C for 20 min
22.	B12 assay agar	51.5	7.2 ± 0.2	Sterilization at 1210C for 20 min
23.	B12 culture agar	37.85	7.0 ± 0.2	Sterilization at 1210C for 20 min
24.	Triple sugar iron agar	65.0	7.4 ± 0.2	Sterilization at 1210C for 20 min