Inside this standard operating procedure, you will learn about the Disposal of Microbial Media culture waste and the Maintenance of Slants / Butt and Culture.
1.0 Objective: To lay down the procedure for the Maintenance and disposal of Microbial media in the Microbiology Laboratory.
2.0 Scope: This Standard Operating procedure for the Maintenance and disposal of Microbial media in the Microbiology Laboratory.
3.0 Responsibility:
Executive Microbiologist/ Microbiologist- for implementation of this SOP.
Head Production:- For overall compliance with this SOP.
4.0 Distribution:
Master Copy – Quality Assurance Department
Control Copy – Quality Assurance Department
Display Copy – Quality Assurance Department
5.0 Procedure:
5.1 Maintenance of Slants / Butt and Culture :
5.1.1 All the received cultures slants/butt are considered a parent culture and labeled as a parent culture indicated by ‘P.’
5.1.2 From each parent’s culture slants/butt streak further into another six fresh pre-incubated slants/butt. Incubate at the specified temperature and period.
5.1.3 Label the slants/butt of each culture as ‘P1 to P6’ and keep P6 slants/butt like a mother culture.
5.1.4 After getting Growth evidence, P1 slants/butt of each culture again streaked into another 5 pre-incubator slants/butt of each culture of P1 and labeled them as ‘P1.1 to P1.5’. Moreover, incubate them at the specified temperature and period.
5.1.5 After getting evidence of growth in ‘P1.1 to P1.5’ slants/butt, further streak again into 5 pre-incubated slants/butt And label them as ‘P1.1.1 to P1.1.5’ for each culture and incubate them at the specified temperature and period.
5.1.6 After getting Growth evidence, use these sub-passage slants/butt ‘P1.1.1 to P1.1.5’ for not more than 15 days.
5.1.7 Repeat step no. 5.1.4, 5.1.5, and 5.1.6 for P2, P3, P4, and P5 slants/butt and maintained the culture for one year, after completion of 1 year again order the culture slants/butt as per requirements.
5.1.8 Never use the sub-passage of culture slants/butt for more than 15 days.
5.1.9 All the cultures slants/butt shall be stored in the refrigerator between 2 to 8°C.
5.1.10 On detecting any contamination in the culture slants/butt, discard that passage or sub-passage and streak again into the fresh pre-incubated slants/butt and assign them as a new passage no.
5.2 Disposal of Microbial Media :
5.2.1 Add a small quantity of a suitable approved disinfectant to each culture slant/butt before their destruction.
5.2.2 Autoclave all the discarded cultures slants/butt at 121°C for 30 minutes at 15 lbs / in2.
5.2.3 To ensure thorough disinfection, all Petri dishes/glassware should be dipped in a 5% Dettol/teepol solution overnight.
5.2.4 Finally Remove them and wash them with Tap water followed by Running purified water.
5.2.5 Dry them in a hot air oven at 180° C for 30 Minutes.
5.2.6 Discard the autoclave culture slants / butt in ETP.
5.2.7 Prepare 70% IPA
5.2.8 After the subculturing, immediately clean the LAF with filtered 70 % IPA and mop the microbiology testing room with any of the approved disinfectants used for that particular date.
6.0 Abbreviations:
SOP- Standard Operating Procedure
IPA -Isopropyl Alcohol
LAF -Laminar Air Flow
QA – Quality Assurance
QC- Quality Control
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