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BAM Media M151a: Trypticase-Peptone-Glucose-Yeast Extract Broth with Trypsin

January 2001

Bacteriological Analytical Manual

M151a Trypticase-Peptone-Glucose-Yeast Extract Broth with Trypsin (TPGYT)

 

Base
 

Trypticase 50 g
Peptone 5 g
Yeast extract 20 g
Dextrose 4 g
Sodium thioglycollate 1 g
Distilled water 1 liter

 

Dissolve solid ingredients of base and dispense 15 ml in 20 x 150 mm tubes or 100 ml in 170 ml prescription bottles. Autoclave tubes 10 min at 121°C and bottles 15 min at 121°C. Final pH, 7.0 ± 0.2. Refrigerate at 5°C. Add trypsin immediately before use.

 

Trypsin solution
 

trypsin (1:250) 1.5 g
Distilled water 100 ml

 

Stir trypsin in water to suspend. Let particles settle and filter-sterilize supernatant through 0.45 µm membrane.

Before use, steam or boil base for 10 min to expel dissolved oxygen. Add 1 ml trypsin to each 15 ml of broth or 6.7 ml trypsin to 100 ml of broth.


Hypertext Source: Bacteriological Analytical Manual, 8th Edition, Revision A, 1998.