Insertion of lux genes into the chromosome of Salmonella enterica Bioluminescence was established in the chromosome of the Salmonella enterica serotypes using plasmid pBEN276. The serotypes were grown to logarithmic phase (OD600 0.6-0.8), washed with 15% cold glycerol solution four times to

make electrocompetent, and stored at -80°C. The serotypes were transformed with plasmid pBEN276 by electroporation using a Gene Pulser II system MM-102 research buy (Bio-Rad). Optimal electroporation conditions for S. Alachua, S. Heidelberg, S. Kentucky, S. Mbandaka, S. Newport and S. Seftenberg were 2.5 kV, 25 μF and 400Ω, and optimal conditions for S. Braenderup, S. Enteritidis, S. Montevideo, S. Schwarzengrund and S. Typhimurium were

1.8 kV, 25 μF and 600Ω. Bacteria were recovered for 1 h at 30°C in SOC media and then spread on LB plates with ampicillin and placed in an incubator at 30°C for approximately 16 h. Ampicillin resistant colonies were picked and cultured in LB broth with arabinose at 30°C for approximately 16 h to induce transposition. The cultures were streaked on LB agar and placed in an incubator at 42°C for approximately 16 h to cure the plasmid. Ten individual colonies were picked {Selleck Anti-cancer Compound Library|Selleck Anticancer Compound Library|Selleck Anti-cancer Compound Library|Selleck Anticancer Compound Library|Selleckchem Anti-cancer Compound Library|Selleckchem Anticancer Compound Library|Selleckchem Anti-cancer Compound Library|Selleckchem Anticancer Compound Library|Anti-cancer Compound Library|Anticancer Compound Library|Anti-cancer Compound Library|Anticancer Compound Library|Anti-cancer Compound Library|Anticancer Compound Library|Anti-cancer Compound Library|Anticancer Compound Library|Anti-cancer Compound Library|Anticancer Compound Library|Anti-cancer Compound Library|Anticancer Compound Library|Anti-cancer Compound Library|Anticancer Compound Library|Anti-cancer Compound Library|Anticancer Compound Library|Anti-cancer Compound Library|Anticancer Compound Library|buy Anti-cancer Compound Library|Anti-cancer Compound Library ic50|Anti-cancer Compound Library price|Anti-cancer Compound Library cost|Anti-cancer Compound Library solubility dmso|Anti-cancer Compound Library purchase|Anti-cancer Compound Library manufacturer|Anti-cancer Compound Library research buy|Anti-cancer Compound Library order|Anti-cancer Compound Library mouse|Anti-cancer Compound Library chemical structure|Anti-cancer Compound Library mw|Anti-cancer Compound Library molecular weight|Anti-cancer Compound Library datasheet|Anti-cancer Compound Library supplier|Anti-cancer Compound Library in vitro|Anti-cancer Compound Library cell line|Anti-cancer Compound Library concentration|Anti-cancer Compound Library nmr|Anti-cancer Compound Library in vivo|Anti-cancer Compound Library clinical trial|Anti-cancer Compound Library cell assay|Anti-cancer Compound Library screening|Anti-cancer Compound Library high throughput|buy Anticancer Compound Library|Anticancer Compound Library ic50|Anticancer Compound Library price|Anticancer Compound Library cost|Anticancer Compound Library solubility dmso|Anticancer Compound Library purchase|Anticancer Compound Library manufacturer|Anticancer Compound Library research buy|Anticancer Compound Library order|Anticancer Compound Library chemical structure|Anticancer Compound Library datasheet|Anticancer Compound Library supplier|Anticancer Compound Library in vitro|Anticancer Compound Library cell line|Anticancer Compound Library concentration|Anticancer Compound Library clinical trial|Anticancer Compound Library cell assay|Anticancer Compound Library screening|Anticancer Compound Library high throughput|Anti-cancer Compound high throughput screening| from this plate and cultured in LB broth at 42°C for approximately 16 h. Bioluminescent colonies were detected using a ChemiImager 5500 imaging system with AlphaEaseFC software (Alpha Innotech) or an IVIS Imaging System 100 Series with Living Image Software v2.50 (Xenogen). Bioluminescent cultures were Torin 2 subcloned in LB broth with ampicillin and placed in an incubator at 30°C for approximately 16 h. No visual evidence of growth confirmed absence of the plasmid. Characterizing the bioluminescent

properties of Salmonella enterica serotypes The bioluminescent Salmonella enterica serotypes were grown overnight in LB broth Rebamipide to reach stationary phase, and bacterial density value (OD600) of each serotype was determined in a 96-well clear-bottomed black cell culture plate (Costar) using ThermoMax spectrometer (Molecular Devices). Following bacterial density measurements, four separate dilution series were prepared for each serotype in 96-well clear-bottomed black cell culture plates. In each plate, the first four columns contained 10 fold dilutions (1.00 × 100 to 1.00 × 10-3), while the remaining eight wells contained doubling dilutions (5.00 × 10-4, 2.50 × 10-4, 1.25 × 10-4, 6.25 × 10-4, 3.13 × 10-5, 1.56 × 10-5, 7.81 × 10-6, 3.91 × 10-6, 1.95 × 10-6, 9.77 × 10-7, 4.88 × 10-7, 2.44 × 10-7). Bioluminescence was measured for 10 s of exposure using an IVIS Imaging System 100 Series, and bioluminescence was quantified using Living Image software v2.50. The last dilution of each series was spread on LB agar to determine the number of viable bacteria.