LB Broth

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	supplement the media with carbenicillin (100 μg/ml), spectinomycin (100 μg/ml), erythromycin (300 μg/ml) for Escherichia coli and (5 μg/ml) for B. anthracis

Protocol tips
supplement the media with carbenicillin (100 μg/ml), spectinomycin (100 μg/ml), erythromycin (300 μg/ml) for Escherichia coli and (5 μg/ml) for B. anthracis

Publication protocol

Bacillus anthracis was cultured in Luria-Bertani (LB) medium (Ausubel, 1993) to prepare cells for electroporation and DNA isolation. Cell lysates and culture supernates for Western blot analysis were obtained from cells cultured in CACO3 medium (Casamino Acids medium [Thorne and Belton, 1957] buffered with 100 mM HEPES [pH 8.0] and 0.8% [wt/vol] sodium bicarbonate) or PA (Phage Assay) medium. Briefly, an overnight culture of B. anthracis grown in LB medium supplemented with appropriate antibiotics and incubated with agitation at 30°C was used to inoculate CACO3 or PA medium comprising 10% of the volume of an Erlenmeyer flask. Cultures were incubated at 37°C with agitation. Cultures in CACO3 medium were incubated in an atmosphere of 5% CO2. Antibiotics were added to media as appropriate: carbenicillin (100 μg/ml), spectinomycin (100 μg/ml), erythromycin (300 μg/ml) for Escherichia coli and (5 μg/ml) for B. anthracis. All chemicals were purchased from Fisher unless otherwise stated.

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