Tetracycline (TCy) and chloramphenicol were purchased from EMD Millipore (Germany). Ampicillin, erythromycin, and kanamycin sulfate were purchased from Bio BasicInc. (USA). Vancomycin was purchased from Calbiochem (USA). Sulfamethoxazole, ciprofloxacin, trimethoprim, and hippuric acid were obtained from Sigma–Aldrich (Germany). Quinine was obtained from Alfa Aesar (USA). CaCl2.2H2O, ZnCl2, KCl, NaCl, and NH4Cl were purchased from Showa (Japan). Polyvinylpyrrolidone (PVP, molecular weight = 58,000) was obtained from Acros Organics (USA). All reagents were reagent grade and were used without further purification. Ultrapure water prepared using a Milli-Q water system (Simplicity, Millipore) was used to prepare aqueous solutions.
Kanamycin sulfate
Manufactured by Bio Basic
Sourced in Canada
Kanamycin sulfate is a broad-spectrum antibiotic used in laboratory settings. It functions as a selective agent for the growth and maintenance of bacteria that are resistant to the antibiotic.
Automatically generated - may contain errors
Lab products found in correlation
T4 dna ligase, by Thermo Fisher Scientific (2 mentions)
Sulfamethoxazole, by Merck Group (1 mentions)
Quinine, by Thermo Fisher Scientific (1 mentions)
Trimethoprim, by Merck Group (1 mentions)
Zncl2, by Resonac (1 mentions)
Sodium chloride (nacl), by Resonac (1 mentions)
Erythromycin, by Bio Basic (1 mentions)
Ciprofloxacin, by Merck Group (1 mentions)
Hippuric acid, by Merck Group (1 mentions)
Chloramphenicol, by Merck Group (1 mentions)
Tetracycline, by Merck Group (1 mentions)
Ampicillin, by Bio Basic (1 mentions)
Ni nta matrix, by Qiagen (1 mentions)
Isopropylthio β dgalactopyranoside iptg, by Merck Group (1 mentions)
Hammerstein casein, by Merck Group (1 mentions)
Lysozyme, by Merck Group (1 mentions)
Wizard genomic dna purification kit, by Promega (1 mentions)
3 protocols using kanamycin sulfate
1
Antimicrobial Agents and Materials
2
Purification and Expression of Recombinant Protein
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The restriction enzymes (XhoI, SalI, and NdeI) were obtained from Jena Bioscience (Jena, Germany), T4 DNA ligase was purchased from Thermo Fisher Scientific (Grand Island, NY). Anti-PF4 antibody (ThermoFisher Scientific Company, mouse monoclonal/IgG2b, clone name 170138, Grand Island, NY) and conjugated anti-mouse antibody (Abcam company, rabbit anti-mouse IgG, polyclonal, ab97046, USA) were used to perform the Western blotting experiments. Immobilized metal affinity chromatography (IMAC) using the Ni-NTA matrix (Qiagen) was employed for purification purposes. The synthetic construct was ordered from Biomatik Company (Biomatik, Ontario, Canada). To alleviate problems regarding E. coli codon bias, the Biomatik's proprietary codon optimization service was exploited. The agarose gel and plasmid extraction kits were purchased from Bioneer (South Korea). Isopropyl beta-D-thiogalactopyranoside (IPTG) and Kanamycin sulfate were acquired from Bio-Basic (Markham, Canada) and Merck (Germany) respectively. pET26b was utilized as the expression vector. BL21 (DE3) and DH5α strains of E. coli were provided by the Diagnostic Laboratory Sciences and Technology Research Center, Shiraz, Iran. The E. coli DH5α strain was employed for plasmid extraction purposes. The E. coli BL21 (DE3) strain was exploited as an expression host.
3
Genomic DNA Purification and Bacterial Culture
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The Wizard® genomic DNA purification kit used was from Promega (Madison, WI, USA). The oligonucleotides used in this work (listed in Supplementary data, Table S1) were Hammerstein casein was provided by Merck (Darmstadt, Germany). Kanamycin sulfate and ampicillin sodium salt USP were purchased from Bio Basic (Markham, Canada Inc.).
Isopropyl-thio-β-D-galactopyranoside (IPTG) and lysozyme were bought from Sigma-Aldrich Co (St Louis, MO, USA).
The culture media used were Luria-Bertani (LB) and yeast extract peptone dextrose (YPD), buffered glycerol-complex (BMGY), and buffered methanol-complex (BMMY). The YPD media were composed of yeast extract, 10 g; peptone, 20 g; and dextrose, 20 g per liter.
The LB and YPD were complemented with 20 g/L agar to make solid media. The BMGY/BMMY medium was composed of yeast extract, 1%; peptone, 2%; biotin, 4×10 -5 % (w/v); and yeast nitrogen base, 1.34% in 100 mM potassium phosphate pH 6 plus glycerol 1%
Isopropyl-thio-β-D-galactopyranoside (IPTG) and lysozyme were bought from Sigma-Aldrich Co (St Louis, MO, USA).
The culture media used were Luria-Bertani (LB) and yeast extract peptone dextrose (YPD), buffered glycerol-complex (BMGY), and buffered methanol-complex (BMMY). The YPD media were composed of yeast extract, 10 g; peptone, 20 g; and dextrose, 20 g per liter.
The LB and YPD were complemented with 20 g/L agar to make solid media. The BMGY/BMMY medium was composed of yeast extract, 1%; peptone, 2%; biotin, 4×10 -5 % (w/v); and yeast nitrogen base, 1.34% in 100 mM potassium phosphate pH 6 plus glycerol 1%
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