BRAIN HEART INFUSION BROTH

Protocol tips

Upstream tips	Protocol tips	Downstream tips
-Store between 10-30°C in a tightly closed container and the prepared medium at 15-25°C.	-Reaction of 3.7% w/v aqueous solution at 25°C. pH : 7.4±0.2. -Cultural characteristics observed after an incubation at 35-37°C for 24-48 hours.

Upstream tips
-Store between 10-30°C in a tightly closed container and the prepared medium at 15-25°C.
Protocol tips
-Reaction of 3.7% w/v aqueous solution at 25°C. pH : 7.4±0.2. -Cultural characteristics observed after an incubation at 35-37°C for 24-48 hours.

Publication protocol

Escherichia coli, used for vector propagation, was grown in Luria-Bertani (LB) broth (Difco) containing 50 μg/ml kanamycin sulfate (Amresco, Solon, OH). S. pyogenes strains were grown in Bacto Todd Hewitt Broth (Becton, Dickinson and Company, Sparks, MD) supplemented with 2% Bacto yeast extract (THY medium) at 37°C or Brain Heart Infusion media (BHI) (Himedia Laboratories, India) at 37°C with the addition of the appropriate antibiotics (Sigma); growth was determined by monitoring the absorbance at 600 nm. Strain SF370SmR is a streptomycin resistant derivative of strain SF370, containing a spontaneous mutation in the rpsL gene. No differences between SF370 and the streptomycin resistant derivative were observed with respect to growth rate or in the various biological assays employed.

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