1.
Scope
·
This procedure is routinely use to check
sterility of prepared media by inoculating in peptone water, thioglycolate
broth and sabouraud agar.
2.
Definition
·
% - Percentage
3. Material
·
Media
prepared in the laboratory
o
MEM growth medium
o
MEM maintenance medium
o
L-Glutamine (3%)
o
Hepes buffer
o
7.5% NaHCO3
o
PBS without Ca ++and Mg++
·
Ready to use media
o
Foetal Bovine Serum
o
Trypsin EDTA
o
PBS with Ca ++and Mg++
4.
Procedure
· Sterility tests of any of these media-
o
Add 2.5ml of each of the above media to two
tubes of peptone broth and thiglycolate broth.
o
Add 1ml of the above media to two tubes of
sabouraud’s agar.
o
Incubate one tube of each at 36°Ċin incubator
and another tube at room temperature.
o
Observe all tubes for 7 days for appearance of
turbidity or fungus in case of sabouraud agar.
o
If there is no turbidity after 7 days, media is
ready for use.
o
If turbidity appears discard media by
autoclaving and prepare fresh media.
1. Safety
conditions
·
Add required amount of media in each tube.
·
Check the sterility tubes everyday and record
it.
2. Documentation
·
Sterility
record for Media File PF-05.
3. References
used to draw SOP
WHO Polio Laboratory Manual
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