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 2-8ÂșC till the new cultures are
received.
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