Listeria monocytogenes EGDe (1/2a, internal number 2964) as well as ΔprfA L. monocytogenes EGDe (1/2a) were part of the collection of bacterial strains at the Institute of Milk Hygiene, Milk Technology and Food Science, University of Veterinary Medicine, Vienna, Austria. All bacterial strains were grown overnight in tryptone soya broth with 0.6% (w/v) yeast extract (TSB-Y; Oxoid, Hampshire, United Kingdom) at 37°C. Enumeration of bacterial suspensions was performed using the plate count method.
Tsb y
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
The TSB-Y is a laboratory equipment product manufactured by Thermo Fisher Scientific. It serves as a growth medium for the cultivation of microorganisms. The TSB-Y provides a nutrient-rich environment to support the proliferation of a variety of bacterial and fungal species.
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Lab products found in correlation
5 protocols using tsb y
1
Listeria monocytogenes Strain Cultivation
2
DNA Isolation from L. monocytogenes
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One milliliter of a L. monocytogenes (strain EGDe 1/2a) or ΔprfA L. monocytogenes (strain EGDe 1/2a [11 ], both part of the collection of bacterial strains at the Institute of Milk Hygiene, Milk Technology and Food Science, University of Veterinary Medicine, Vienna, Austria) overnight culture (grown in tryptone soya broth with 0.6% (w/v) yeast extract (TSB-Y; Oxoid, Hampshire, UK) at 37 °C) was used for DNA isolation. The DNA concentration was measured with the Qubit dsDNA Broad Range Kit (Fisher Scientific, Vienna, Austria). The copy number of the single-copy prfA gene was calculated using the molecular weight (1 ng of DNA equals 3.1 × 105 copies of the genome).
3
Bacterial Strains Maintenance and Cultivation
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The eae-positive enteropathogenic E. coli O8:H14 strain (EPEC) obtained from AGES (Vienna, Austria) and community-associated methicillin-resistant S. aureus (CA-MRSA) strain NRS384 (also known as USA300-0114) obtained from the Network on Antimicrobial Resistance in Staphylococcus aureus (NARSA) collection were maintained at −80 °C using MicroBankTM technology (Pro-Lab Diagnostics, Richmont Hill, ON, Canada). All bacterial strains were grown overnight in tryptone soya broth with 0.6% (w/v) yeast extract (TSB-Y; Oxoid, Basingstoke, Hampshire, UK) at 37 °C.
4
Cultivation of L. monocytogenes Strains
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L. monocytogenes EGDe (1/2a, internal number 2964) as well as ΔprfA L. monocytogenes EGDe (1/2a) were part of the collection of bacterial strains at the Institute of Milk Hygiene, Milk Technology and Food Science, University of Veterinary Medicine, Vienna, Austria. All bacterial strains were grown overnight in tryptone soya broth with 0.6 % (w/v) yeast extract (TSB-Y; Oxoid, Hampshire, UK) at 37 °C. Enumeration of bacterial suspensions was performed using the plate count method.
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L. monocytogenes EGDe (1/2a, internal number 2964) as well as ΔprfA L. monocytogenes EGDe (1/2a) were part of the collection of bacterial strains at the Institute of Milk Hygiene, Milk Technology and Food Science, University of Veterinary Medicine, Vienna, Austria. All bacterial strains were grown overnight in tryptone soya broth with 0.6 % (w/v) yeast extract (TSB-Y; Oxoid, Hampshire, UK) at 37 °C. Enumeration of bacterial suspensions was performed using the plate count method.
5
Cryogenic Preservation of Listeria Strains
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All strains (Table 1) were stored in 2 ml cryovials with a 7% dimethyl sulfoxide (DMSO) at -80 o C. L. monocytogenes 10403S and EGD-e were cultured onto Brain Heart Infusion (BHI) agar (LABM, Lancashire UK) and L. monocytogenes LO28 onto Tryptic Soy Broth (Oxoid, UK) supplemented with 5% yeast extract (TSBY; Oxoid, UK) and incubated at 37 o C overnight.
Three colonies from each plate were transferred, with an inoculation loop, into BHI and TSBY broth respectively in 10 ml Sterilin polystyrene tubes and incubated at 37 o C with shaking (150 rpm) for 18 h. These overnight cultures were used to inoculate 20 ml cultures of the corresponding media (1% inoculum) in 250 ml conical flasks which then were subsequently incubated overnight at 37 o C with shaking at (150 rpm) for 18 h.
Three colonies from each plate were transferred, with an inoculation loop, into BHI and TSBY broth respectively in 10 ml Sterilin polystyrene tubes and incubated at 37 o C with shaking (150 rpm) for 18 h. These overnight cultures were used to inoculate 20 ml cultures of the corresponding media (1% inoculum) in 250 ml conical flasks which then were subsequently incubated overnight at 37 o C with shaking at (150 rpm) for 18 h.
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