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M151a
Trypticase-Peptone-Glucose-Yeast Extract Broth with Trypsin
(TPGYT)
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 (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 updated by rim/cjm 2005-OCT-21