BAM Media M149: Triple Sugar Iron Agar (TSI)

January 2001

Bacteriological Analytical Manual

Triple Sugar Iron Agar (TSI)

Medium 1

Medium 2

Polypeptone20 gBeef extract3 g
NaCl5 gYeast extract3 g
Lactose10 gPeptone15 g
Sucrose10 gProteose peptone5 g
Glucose1 gGlucose1 g
Fe(NH4)2(SO4)2•6H2O0.2 gLactose10 g
Na2S2O30.2 gSucrose10 g
Phenol red0.025 gFeSO40.2 g
Agar13 gNaCl5 g
Distilled water1 literNa2S2O30.3 g
 Phenol red0.024 g
Agar12 g
Distilled water1 liter

These two media are interchangeable for general use.

Suspend ingredients of Medium 1 in distilled water, mix thoroughly, and heat with occasional agitation. Boil about 1 min to dissolve ingredients. Fill 16 × 150 mm tubes 1/3 full and cap or plug to maintain aerobic conditions. Autoclave Medium 1 for 15 min at 118°C.

Prepare Medium 2 in the same manner as Medium 1, except autoclave 15 min at 121°C. Before the media solidify, incline tubes to obtain 4-5 cm slant and 2-3 cm butt. Final pH, 7.3 ± 0.2 for Medium 1 and 7.4 ± 0.2 for Medium 2.

For use with halophilic Vibrio spp., add NaCl to a final concentration of 2-3%.

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

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