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Luria bertani medium

Manufactured by Nacalai Tesque
Sourced in Japan

Luria-Bertani (LB) medium is a widely used nutrient-rich growth medium for cultivating bacteria. It provides essential nutrients, such as amino acids, vitamins, and minerals, to support the growth and proliferation of various bacterial species. LB medium is commonly used in molecular biology, microbiology, and biotechnology applications.

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2 protocols using luria bertani medium

1

Streptomycin-Resistant E. coli Strain S-17 Characterization

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streptomycin-resistant green fluorescent protein (GFP)-expressing E. coli strain S-17 (S-17-GFP) was generated as described previously75 (link). S-17-GFP were cultured in Luria-Bertani medium (Nacalai Tesque) with 50 μg/mL streptomycin (Wako) overnight, then bacteria were transferred into LB medium with 1 mM isopropyl β-D-1-thiogalactopyranoside (Wako) and incubated at 37 °C for an hour. After incubation, bacteria were resuspended in 800 μL of 4% paraformaldehyde at 4 °C for 3 h. Then bacteria were resuspended into PBS and measured OD 600 by spectrophotometer. A total of 1 × 107 cells (assuming OD 600 1.0 = 5 × 108 cell/mL) bacteria were transferred to a new tube and resuspended and incubated with 10 μg rGP2 or PBS at 4 °C overnight. GP2-bound bacteria were detected by using a combination of anti-mGP2 antibody diluted 1:100 and PE-conjugated anti-rat IgG2a antibody diluted 1:100. Flow cytometry analysis was performed with a FACSCanto II instrument or ATTUNE Next. Gating strategies for the bacterial flow cytometry were shown in Supplementary Fig. 18.
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2

Streptococcus pyogenes Culturing and Manipulation

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Streptococcus pyogenes M1T1 strain 5448 (accession: CP008776.1) was isolated from a patient with toxic shock syndrome and necrotizing fasciitis that is genetically representative of a globally disseminated clone associated with invasive S. pyogenes infections (Kansal et al., 2000 (link)). S. pyogenes strains were grown at 37°C in a screw-cap glass tube (Pyrex; Iwaki Glass, Tokyo, Japan) filled with Todd-Hewitt broth (BD Biosciences, San Jose, CA, USA) supplemented with 0.2% yeast extract (BD Bioscience) (THY broth) in an ambient atmosphere and standing cultures. To obtain cultures for experiments and observe pH change, overnight cultures of S.pyogenes were back diluted 1:50 into fresh THY broth or phenol red broth (Sigma Aldrich, St Louis, MO, USA) supplemented with 30 mM arginine. CFUs were determined by plating diluted samples on THY blood agar.
Escherichia coli strain XL-10 Gold (Agilent Technologies, Santa Clara, CA, USA) was used as a host for derivatives of plasmids pSET4s (Takamatsu et al., 2001 (link)) and pQE30 (Qiagen, Hilden, Germany). E. coli strains were cultured in Luria-Bertani medium (Nacalai Tesque, Kyoto, Japan) at 37°C with agitation. For selection and maintenance of strains, antibiotics were added to the medium at the following concentrations: spectinomycin, 100 μg/mL for S. pyogenes and E. coli: carbenicillin, 100 μg/mL for E. coli.
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