Luria bertani broth

Total bacterial DNA from overnight culture in Luria-Bertani Broth (Biocorp, Warszawa, Poland) was extracted using commercial Genomic Mini kit (A&A Biotechnology, Gdynia, Poland) according to the manufacturer’s protocol.

Bacteria were grown in Luria-Bertani Broth (Biocorp, Warszawa, Poland) for 16 h at 28 °C. Then 1 mL of bacterial suspension was centrifuged at 10,000 rpm, for 10 min at room temperature. Extraction of genomic DNA was carried out using mini-columns from Genomic Mini kit (A&A Biotechnology, Gdańsk, Poland), according to the manufacturer’s protocol.

A sterile microloop was used to take a few colonies of a pure culture cultivated on Luria-Bertani Broth (Biocorp, Warszawa, Poland) for 18 to 24 h. A bacterial suspension was adjusted to McFarland Turbidity Standard of 0.5–0.63 in 3 mL of a 0.45% sodium chloride solution with a VITEK^® 2 DensiChek (bioMérieux, Warszawa, Poland). The GN card was put on the cassette and placed in the instrument. The time interval between suspension preparation and card filling was less than 30 min to avoid changes in turbidity. Cards were incubated at 35.5 ± 1 °C. Each card was removed from the incubator every 15 min and automatically subjected for colorimetric readings. Results were read after 10 h – 18 h of incubation.