Tryptic Soy Broth for microbiology

Protocol tips

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	-Anticoagulants such as sodium polyanetholesulfonate (81305) or sodium citrate (71635) may be added to the broth prior to sterilisation. 5 to 10 ml of blood may be added to 50 ml of medium. -Final pH 7.3 +/- 0.2 at 25°C

Protocol tips
-Anticoagulants such as sodium polyanetholesulfonate (81305) or sodium citrate (71635) may be added to the broth prior to sterilisation. 5 to 10 ml of blood may be added to 50 ml of medium. -Final pH 7.3 +/- 0.2 at 25°C

Publication protocol

S. typhi was grown on TSB for 12 hours at 37°C. The cells were collected in the mid-log phase of the culture (O.D. ~ 0.6, approximately corresponding to 5×108 CFU/mL). The correlation between optical density and CFU/mL was obtained via serial dilution and plating method. The cells were spiked in sterile blood culture media and diluted serially to obtain 500, 400, 300, 200, 100, 50, 10 and 5 CFU/mL. All serial dilutions were carried out using sterile blood culture media. The mixtures were vortexed for 30 seconds for homogenous mixing. The sterile blood culture media without spiking of S. typhi was used as negative control. One milliliter solution of each concentration was used for obtaining the first standardization curve. The spiked blood cultures were incubated for 4 hours at 37°C for growth of the S. typhi cells. Post incubation, one milliliter of a solution of each concentration was used for obtaining the second standardization curve. To obtain the concentration of S. typhi for pre- and post-four-hour incubation, 20 μL of each dilution was plated on MacConkey® agar plates followed by colony counts after 24 hours.

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Manufacturer protocol

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