Preparation of Salmonella typhimurium Culture

Abstract

Source: Gutiérrez, S., et. al., Isolation of Salmonella typhimurium-containing Phagosomes from Macrophages. J. Vis. Exp. (2017)

This video demonstrates the culturing and preparation of Salmonella typhimurium to obtain a clean, metabolically active bacterial suspension suitable for experimental use.

Protocol

All the steps involving the use of pathogenic Salmonella typhimurium must be carried out in a BSL-2 (Biosafety Level 2) or higher biological security level facility.

1. Culturing Salmonella typhimurium

1. Inoculate Salmonella typhimurium (SL1344 strain) from a single bacterial colony into 5 mL of brain heart infusion (BHI) broth using a bacterial loop.
2. Incubate the bacterial suspension in an incubator at 37 °C overnight with shaking.
3. On the following day, transfer 1 mL of the bacterial suspension into 19 mL of BHI broth in a conical flask and incubate at 37 °C in an incubator with shaking.
4. Monitor the S.

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
MgCl₂	Carl Roth	A537.4
Sucrose	Carl Roth	4621.1
Mannitol	Carl Roth	4175.1
SmartSpec 3000 Spectrophotometer	Bio-Rad	170-2501
Bacterial loop (10µl)	Sarstedt	86.1562.010
Salmonella enterica serovar Typhimurium SL1344	Leibniz Institute DSMZ-German collection of Microorganisms and Cell Cultures
PBS (Phosphate-buffered saline)	Biochrom	L1825