procedure maintenance of microbial culture.

      1.0       Objective

     To lay down a procedure  maintenance of microbial culture.

      2.0        Responsibility

      Microbiologist/QC-Executive.

      3.0        Accountability

      Head-QC executive

4.0      Procedure 

4.1        Sub culturing of microbial culture.

    

      4.1.1    Receive the culture (in powder basis), and transfer it in to broth nutrient and incubate it at temp.37⁰c for                    24 hour.

      4.1.2    Then prepare serial dilution like 10^-1, 10^-2, and 10^-3------10^-5.Transfer 1ml of solution in to serial dilution.

      4.1.3     Then transfer 1ml of sample in Petri plates and pour 15-20ml of nutrient agar in Petri plates.

      4.1.4      Perform Identification test for each received microbial culture as per log book.

      4.1.5      Prepare the slants of required media as per SOP.

   4.1.6      Aseptically streak the colonies in slant and incubate these slants at temp. According to growth.

   4.1.7      Aseptically streak one loop mother culture slant tubes.

   4.1.8      Keep one slant as stock culture and another slant as working culture.

      4.1.9    Incubate the slants for enumeration of bacteria, at 30° to 35° C for 48 hours and at 20° to 25° C for                     enumeration of fungi, for 72 hours.

4.1.10     After incubation period observe the growth on slants, store the slants at 2-8°C in the refrigerator.

4.1.11     Record the culture identification results in log book.

4.1.12     Subculture these slant.

4.1.13    Maintain the record of subculturing in culture record

4.1.14   Perform Gram staining as per SOP of each culture generation during sub culturing.             

      4.1.15   Destroy the old culture after subculturing but preserve the mother culture at 2-8ºC till the new cultures                    are received.