All chemicals used in this work were of analytical grade. IPTG, ZnSO4, kanamycin, biotin and the buffer components were purchased from Sigma–Aldrich (St. Louis, MO, USA). Ampicillin and Nutrient Agar were purchased from Merck Millipore (Darmstadt, Germany). SDS buffer and the SDS-PAGE gels were purchased from Bio-Rad (Hercules, CA, USA). LB media was purchased from Carl Roth (Karlsruhe, Germany) and the protein marker for SDS PAGE from VWR International (Radnor, PA, USA). Bottled CO2 (purity 99.5%) was derived from Messer (Bad Soden, Germany).
Lb media
Manufactured by Carl Roth
Sourced in Germany
LB media is a widely used nutrient-rich growth medium for the cultivation of various bacterial strains in laboratories. It provides essential nutrients, such as amino acids, carbohydrates, and vitamins, to support the growth and proliferation of bacterial cells.
Automatically generated - may contain errors
Lab products found in correlation
Nutrient agar, by Merck Group (1 mentions)
Kanamycin, by Merck Group (1 mentions)
Znso4, by Merck Group (1 mentions)
Isopropyl β d 1 thiogalactopyranoside (iptg), by Merck Group (1 mentions)
Sds page gel, by Bio-Rad (1 mentions)
Ampicillin, by Merck Group (1 mentions)
Genejet genomic dna purification kit, by Thermo Fisher Scientific (1 mentions)
Glycerine, by Carl Roth (1 mentions)
Luna universal qpcr master mix, by New England Biolabs (1 mentions)
Nanophotometer, by Implen (1 mentions)
Chloroform, by Thermo Fisher Scientific (1 mentions)
Gentamicin, by Thermo Fisher Scientific (1 mentions)
Meropenem, by Cayman Chemical (1 mentions)
Ham s f12 medium, by GE Healthcare (1 mentions)
Polymyxin b, by Merck Group (1 mentions)
Macconkey agar, by Carl Roth (1 mentions)
Phosphate buffered saline (pbs), by Capricorn (1 mentions)
Methanol, by Merck Group (1 mentions)
Lipopolysaccharides (lps), by Enzo Life Sciences (1 mentions)
Sodium acetate, by Carl Roth (1 mentions)
4 protocols using lb media
1
Chemicals and Materials for Biochemical Experiments
2
Quantitative Detection of Pseudomonas aeruginosa
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All chemicals, unless stated otherwise, were purchased from Sigma-Aldrich, subsidiary of Merck (Darmstadt, Germany). LB media, agar agar, diethylamine, glycerine, PD-tips, and sodium hypochlorite were purchased from Carl Roth (Karlsruhe, Germany). Polyglycerol-3-glycidyl ether (CL9) was obtained from ipox chemicals (Laupheim, Germany). All oligonucleotides were synthesized by Eurofins Genomics (Ebersberg, Germany), purchased in lyophilized form and dissolved in nuclease-free water. Primers for detection of the P. aeruginosa regA gene were designed in house. For genomic DNA extraction, the extraction kit GeneJET Genomic DNA Purification Kit by Thermo Scientific (Waltham, USA) was used. Photometric measurements were performed on a NanoPhotometer by Implen (Munich, Germany). For qPCR, the Luna® Universal qPCR Master Mix by New England Biolabs (Ipswitch, USA) was used. For all experiments, ultrapure water was used, unless stated otherwise. Nuclease-free water was purchased from Invitrogen AG (Carlsbad, USA). A bacterial isolate of Pseudomonas aeruginosa (ATCC27853) was received from the Bavarian Health and Food Safety Authority. Viable bacteria were handled in laboratories approved for biosafety level 2.
3
Bacterial membrane characterization protocol
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LB media, MacConkey agar, Colistin sulfate and sodium acetate were acquired from Carl Roth GmbH & Co. KG (Karlsruhe, Germany). PBS was obtained from Capricorn Scientific GmbH (Ebsdorfergrund, Germany). Ham's F12 medium was purchased from GE Healthcare Europe (Freiburg, Germany). Polymyxin B was supplied by Merck Millipore (Billerica, USA). Gentamicin was obtained from Gibco Life Technologies (Carlsbad, USA) and Meropenem from Cayman Chemicals (Ann Arbor, USA). Methanol was acquired from Merck KGaA (Darmstadt, Germany). Chloroform was purchased from Acros Organics (Geel, Belgium). LPS (Salmonella minnesota R595, TLR grade) was supplied by Enzo Life Sciences (Lausen, Austria). All lipids were obtained from Avanti® Polar Lipids Inc. (Birmingham, USA). All other applied chemicals were of analytical grade and acquired from commercial sources.
4
Staphylococcus Carriage in Kenyan Shelter Dogs
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Diagnostic specimens from all dogs (N = 167) housed at the animal shelter were collected on the 27th June 2015 in Nairobi, Kenya by veterinarians using the TRANSWAB® Amies swabs. Nasal specimens were collected by inserting the swabs up to 10 cm into the nasal cavity of the animals, which were restrained without sedation for the procedure. Wound infections were also swabbed when present. The Staphylococcaceae strains were isolated at the laboratories of the International Livestock Research Institute (ILRI) using standard methods without selective enrichment (44 ). Briefly, each swab was streaked on Baird Parker Agar (Carl Roth) plates and incubated at 37°C overnight. One to two suspicious colonies per plate were picked, expanded in LB media (Carl Roth), and stored as glycerol stocks at −80°C before being shipped for further characterization to the Institute of Veterinary Bacteriology (IVB) in Switzerland. Strains were streaked onto Trypticase Soy Agar (TSA-B; Becton Dickinson) with 5% sheep blood (TSA-B; Becton Dickinson) and incubated at 37°C overnight. Species designation was assigned using MALDI-TOF MS (Bruker) as previously reported (30 (link)) followed by metabolic phenotyping (see below). Data on strains investigated in this study are provided in Dataset S1.
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