Subculturing of Master Cultures | Maintenance of Microbiological Cultures |
1.0 OBJECTIVE :
- To provide a procedure for Culture Transfer and Maintenance of Microbial Cultures.
2.0 SCOPE :
- This procedure applies only for the cultures used in Microbiology Section
3.0 RESPONSIBILITY :
- Microbiologist
4.0 ACCOUNTABILITY:
- Head of Department
5.0 PROCEDURE :
5.1 MAINTENANCE OF CULTURES :
- Check the purity of standard stock culture.
- Prepare two master cultures from standard stock culture.
- One of the two master cultures must be used for preparation of working cultures in first six months.
- The other master culture is used for preparation of working cultures in next six months.
- Prepare monthly working cultures from master culture.
- From the monthly working culture, prepare the routine cultures.
- Use one routine culture for one week.
- All the cultures must be stored in refrigerator at 2°C to 8°C.
- The second master culture is used to prepare working cultures of the next six months further, follow the same steps as above.
5.2 PROCEDURE :
- Prepare SCDA for bacterial culture and SCA for fungal culture as per the Preparation and Destruction of Media for Microbiological Analysis – SOP
- Prepare the slant and keep in incubator.
- Strick the bacterial culture for SCDA slant and fungal culture for SCA slant.
- Place the bacterial slant at 30°C to 35°C for 24 hrs and fungal slants at 20°C to 25°C for 3-5 days.
- After completion of incubation period, store these subculture slants into refrigerator.
- Each slant have a proper label having the details of name of culture , Batch Number of culture, use date, use before date etc.
5.3 FREQUENCY :
- Monthly
- For Vitamin assay culture: Daily culture transfer or sub culturing on fresh media, as per need (on culture agar & inoculums medium).
- The maintenance media required for subculturing of the different microbiological cultures are as follows:
| Sr. No. | Name of cultures | Culture code no as per IP | Maintenance medium |
| 1 | Escherichia coli | ATCC 8739 | Soyabean Casein Digest Agar |
| 2 | Salmonella abony | NCTC 6017 | Soyabean Casein Digest Agar |
| 3 | Pseudomonas aeruginosa | ATCC 9027 | Soyabean Casein Digest Agar |
| 4 | Staphylococcus aureus | ATCC 6538 | Soyabean Casein Digest Agar |
| 5 | Candida albicans | ATCC 10231 | Sabouraud Chloramphenicol Agar |
| 6 | Bacillus subtilis | ATCC 6633 | Soyabean Casein Digest Agar |
| 7 | Escherichia coli( mutant) | ATCC 11105 | Soyabean Casein Digest Agar |
| 8 | Aspergillus brassiliances | ATCC 16404 | Sabouraud Chloramphenicol Agar |
| 9 | Shigella boydii | ATCC 8700 | Soyabean Casein Digest Agar |
6.0 ABBREVIATIONS:
| Abbreviation | Expanded form |
| SCDA | Soybean Casein Digest Agar |
| SCA | Sabouraud Chlormaphenicol Agar |
| SOP | Standard Operating Procedure |
| ATCC | American Type Culture Collection |
| IP | Indian Pharmacopoeia |
7.0 ANNEXURES:
| Annex. No. | Title |
| 01 | Subculturing Of Master and Maintenance Of Microbiological Cultures |
8.0 SOP REFERENCES
- Indian Pharmacopoeia
END OF THE SOP
ANNEXURES :
Annex. No. 01 Subculturing of Master and Maintenance of Microbiological Cultures
