Luria Bertani Broth, Miller (Miller Luria Bertani Broth)

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Store below 30°C in tightly closed container and the prepared medium at 2 - 8°C.	-Reaction of 2.5% w/v aqueous solution at 25°C. pH : 7.5±0.2

Upstream tips
Store below 30°C in tightly closed container and the prepared medium at 2 - 8°C.
Protocol tips
-Reaction of 2.5% w/v aqueous solution at 25°C. pH : 7.5±0.2

Publication protocol

Escherichia coli XL1-Blue and BL21 (DE3) pLysS were used for cloning and expression of the recombinant proteins, respectively. E. coli strains were cultured in Luria Bertani (LB) broth (Himedia) and M. abscessus subsp. massiliense (Cardoso et al., 2008) was cultured in Mueller Hinton broth or agar at 37°C under 180 rpm shaking. For growth in different iron concentrations, the minimal medium contained 3.6 mM of KH2PO4, 2.0 mM of MgSO4 7H2O, 6% (v/v) of glycerol, 30 mM of l-asparagine, 0.006 mM of ZnSO4, and 0.05% (v/v) of Tween 80, pH 6.8 in iron free conditions. For low iron conditions, media was supplemented with 1.25 mM of deferoxamine mesylate (DFO). In high iron conditions, media was supplemented with 50, 150, 300, or 450 μM of FeCl3. Minimal media without supplementation (iron or DFO) contains enough iron concentration to sustain M. abscessus subsp. massiliense growth, and thus this condition was used as normalizer. E. coli transformants were selected in medium supplemented with the antibiotics kanamycin (KAN) and chloramphenicol (CAM) at 20 μg/ml..

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