Culture Suspension Technique:-
A suspension culture, also known as a cell suspension, is a type of cell culture in which single cells or small aggregates of cells are allowed to function and multiply in an agitated growth medium.
1.0 OBJECTIVE:
To provide a guideline for the Preparation of culture suspension in Microbiology Lab
2.0 SCOPE:
2.1 The procedure is applicable to all personnel working in Microbiology Lab.
3.0 RESPONSIBILITY:
3.1 Microbiologists/Personnel Working in Microbiology Lab
3.1.1 To follow the procedure while performing the Microbiological activities.
3.1.2 To train the new employees during on job training
3.2 Head-Microbiology
3.2.1 To ensure compliance to the procedures
3.2.2 To ensure all the personnel are trained and adhere to the procedures
4.0 ACCOUNTABILITY :
4.1 Head Quality Assurance.
5.0 Procedure:-
Preparation of Microbial Inoculums:
list of Micro organism used in culture suspension Activity.
- Prepare the culture slant as per SOP for Culture maintenance.
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Take a loop full of culture from the working culture slant and inoculate into 10 mL Soybean Casein Digest Medium with specified organisms.
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Incubate for about 24 – 48 hrs. at 30 – 35 ºC for growth of organism.
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After incubation period, take 1 mL from broth into 10 mL normal saline solution (0.9% w/v) and vortex it to mix uniformly. Label this solution as 10-1.
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Take 1 mL from 10-1 to next dilution of 9 mL of saline solution and label as 10-2 and carry out serial dilutions up to 10-9 as per below diagram.
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Pour 1 mL saline from each slant and pour with SCDA. Incubate it and observe that on which slant to obtain 100 cfu colony.
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Take that no. of tube for GPT test. Store that suspension at 2 – 8 ºC temp.
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Prepare Soybean Casein Digest Agar/Saboured Dextrose Agar as per respective SOP Media Preparation.
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Take 1 mL of each dilution into sterilized petri plates in duplicate and pour 15 – 20 mL of Soyabean Casein Digest Agar.
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Swirl the plates properly in clockwise and anticlockwise directions to mix the content properly, in such away to avoid bubble formation.
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Allow the plates to solidify, incubate the plates at 30 – 35 °C for 24 – 48 hrs to Bacterial & 20 – 25 °C . for 3-5 days to fungi.
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Count the number of colonies in each plate and select the dilution with 10 – 100 cfu/ mL.
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Store the Qualified dilution of 10 – 100 cfu/ mL at 2 – 8 °C
Culture Suspension Technique
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