Bacterial cell culture media Bacillus anthracis

Bacterial culture is a process of letting bacteria multiply in a controlled fashion (temperature, humidity, oxygen content or shaking), in a predetermined culture medium (antibiotic resistance to obtain homogenous clones). It is an important step, especially during cloning, as a single cell can be grown homogeneously (on semi-solid or in liquid conditions) to obtain colonies. As mentioned, bacteria can be cultured in broth cultures (Luria broth or LB) or Petri dishes (Agar plates). A specific antibiotic can be added to the broth or agar plates in order to grow bacteria which have the gene insert conferring its resistance to that antibiotic. Following points are necessary to consider for optimal growth conditions: 1. In general, most bacteria grow well at 37C, but there are some strains which require growth temperatures between 25-30C. 2. It is ideal in broth cultures to fill the flask to ⅓ or less of the total flask volume for optimal aerobic growth. 3. Shaking speeds between 140-180 rpm are appropriate to ensure aeration and that the cells are surrounded by fresh media, and do not settle.

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Protocol tips
Brain-heart infusion (BHI) and nutrient broth yeast extract (NBY) media were from Difco (Difco, Detroit, MI) and Radioselectan was from Schering (Renografin 76%; Schering, Lys Lez-Lannoy, France). Sequencing grade modified trypsin was from Promega (Promega, Madison, WI, USA) and endoproteinase Glu-C from Sigma (Sigma Chemical Co.). Synthetic peptides were synthesized by Bachem (Burgdorf, Switzerland). Tween-80, lecithin, histidine and tryptic soy broth (TSB) were from Sigma-Aldrich (Sigma Chemical Co.). Ultrapure water was from a Milli-Qplus 185 purifier (Millipore, Bedford, MA), HPLC-grade acetonitrile was from SDS (Peypin, France), and analytical-grade formic acid and trifluoroacetic acid (TFA) were from Sigma-Aldrich (Sigma Chemical Co.).
LB Broth

Thermo Fisher Scientific

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
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