Escherichia coli strains were grown at 37°C in LB (Luria Broth, Difco Laboratories) [22 ]. E. coli TG1 was used as the host for plasmid constructions. E. coli HB101 (pRK24) was used for mating experiments. The following antibiotics (Eurobio) were added to the culture medium: kanamycin (40 mg L-1), ampicillin (100 mg L-1), chloramphenicol (12.5 or 25 mg L-1). C. difficile strains, 630Δerm [23 (link)],NF2184 (the cwp84 mutant strain, [ErR]) [9 (link)] and the NF2184 containing the pMTL960Ωcwp84 or the pVP75 plasmids, were grown at 37°C in Brain Heart Infusion (BHI) or BHIS+ glucose (BHI supplemented with Yeast Extract (5mg.ml-1, Difco), cysteine (0.1%; Fisher) and glucose (0.1 M; Sigma)). The following antibiotics were added to the culture medium: thiamphenicol (15 mg L-1, MP Biomedical), supplement (composed of 250 mg L-1 of D-cycloserine and of 8 mg mL-1 of cefoxitin, Oxoid), and erythromycin (5 mg L-1, sigma-aldrich). C. difficile strains were grown in anaerobic condition (90% N2, 5% CO2 and 5% H2; Messer) in anaerobic chamber. The CD1064, P30, 4684/08, 3457, 95–1078, R20291, VPI11186, CD196, 79685, CD4, IT1106 and 95–1578 come from our lab collection [19 (link)]. Ribotype analysis was performed by “CNR Bactéries anaérobies et botulisme (Clostridium difficile)—Laboratoire Associé” (Hopital Saint-Antoine, Paris) using the method described by Stubbs et al. [24 (link)].
Ampicillin
Manufactured by Eurobio Scientific
Ampicillin is a broad-spectrum antibiotic that belongs to the penicillin class. It is commonly used in laboratory settings for the selection and growth of bacterial cultures.
Automatically generated - may contain errors
Lab products found in correlation
Yeast extract, by Solabia (3 mentions)
Lb medium, by Merck Group (3 mentions)
Glycerol, by Euromedex (3 mentions)
Bacto tryptone, by Solabia (3 mentions)
Isopropyl β d thiogalactopyranoside, by Euromedex (3 mentions)
Complete mini edta free protease inhibitor, by Roche (2 mentions)
Cysteine, by Thermo Fisher Scientific (1 mentions)
Kanamycin, by Eurobio Scientific (1 mentions)
Cefoxitin, by Thermo Fisher Scientific (1 mentions)
Thiamphenicol, by MP Biomedicals (1 mentions)
D cycloserine, by Thermo Fisher Scientific (1 mentions)
Glucose, by Merck Group (1 mentions)
Erythromycin, by Merck Group (1 mentions)
Hexafluoroisopropanol, by Merck Group (1 mentions)
Sodium ascorbate, by Thermo Fisher Scientific (1 mentions)
Acetonitrile, by Avantor (1 mentions)
Methanol, by Avantor (1 mentions)
N n n n n pentamethyl diethylenetriamine pmdeta, by Merck Group (1 mentions)
Trizma, by Merck Group (1 mentions)
Ethanol, by Avantor (1 mentions)
4 protocols using ampicillin
1
Cultivation and Genetic Manipulation of E. coli and C. difficile
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Escherichia coli strains were grown at 37°C in LB (Luria Broth, Difco Laboratories) [22 ]. E. coli TG1 was used as the host for plasmid constructions. E. coli HB101 (pRK24) was used for mating experiments. The following antibiotics (Eurobio) were added to the culture medium: kanamycin (40 mg L-1), ampicillin (100 mg L-1), chloramphenicol (12.5 or 25 mg L-1). C. difficile strains, 630Δerm [23 (link)],NF2184 (the cwp84 mutant strain, [ErR]) [9 (link)] and the NF2184 containing the pMTL960Ωcwp84 or the pVP75 plasmids, were grown at 37°C in Brain Heart Infusion (BHI) or BHIS+ glucose (BHI supplemented with Yeast Extract (5mg.ml-1, Difco), cysteine (0.1%; Fisher) and glucose (0.1 M; Sigma)). The following antibiotics were added to the culture medium: thiamphenicol (15 mg L-1, MP Biomedical), supplement (composed of 250 mg L-1 of D-cycloserine and of 8 mg mL-1 of cefoxitin, Oxoid), and erythromycin (5 mg L-1, sigma-aldrich). C. difficile strains were grown in anaerobic condition (90% N2, 5% CO2 and 5% H2; Messer) in anaerobic chamber. The CD1064, P30, 4684/08, 3457, 95–1078, R20291, VPI11186, CD196, 79685, CD4, IT1106 and 95–1578 come from our lab collection [19 (link)]. Ribotype analysis was performed by “CNR Bactéries anaérobies et botulisme (Clostridium difficile)—Laboratoire Associé” (Hopital Saint-Antoine, Paris) using the method described by Stubbs et al. [24 (link)].
+ Expand
Escherichia coli strains were grown at 37°C in LB (Luria Broth, Difco Laboratories) [22 ]. E. coli TG1 was used as the host for plasmid constructions. E. coli HB101 (pRK24) was used for mating experiments. The following antibiotics (Eurobio) were added to the culture medium: kanamycin (40 mg L-1), ampicillin (100 mg L-1), chloramphenicol (12.5 or 25 mg L-1). C. difficile strains, 630Δerm [23 (link)],NF2184 (the cwp84 mutant strain, [ErR]) [9 (link)] and the NF2184 containing the pMTL960Ωcwp84 or the pVP75 plasmids, were grown at 37°C in Brain Heart Infusion (BHI) or BHIS+ glucose (BHI supplemented with Yeast Extract (5mg.ml-1, Difco), cysteine (0.1%; Fisher) and glucose (0.1 M; Sigma)). The following antibiotics were added to the culture medium: thiamphenicol (15 mg L-1, MP Biomedical), supplement (composed of 250 mg L-1 of D-cycloserine and of 8 mg mL-1 of cefoxitin, Oxoid), and erythromycin (5 mg L-1, sigma-aldrich). C. difficile strains were grown in anaerobic condition (90% N2, 5% CO2 and 5% H2; Messer) in anaerobic chamber. The CD1064, P30, 4684/08, 3457, 95–1078, R20291, VPI11186, CD196, 79685, CD4, IT1106 and 95–1578 come from our lab collection [19 (link)]. Ribotype analysis was performed by “CNR Bactéries anaérobies et botulisme (Clostridium difficile)—Laboratoire Associé” (Hopital Saint-Antoine, Paris) using the method described by Stubbs et al. [24 (link)].
2
Clickable Glycoconjugate Synthesis Protocol
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Bacto-tryptone and yeast extract were obtained from Biokar Diagnostics (FR). LB medium was purchased from Sigma-Aldrich (FR). Ampicillin was purchased from Eurobio (FR). Glycerol and isopropyl β-D-thiogalactopyranoside (IPTG) were purchased from Euromedex (FR).
Glycidyl propargyl ether was obtained from Sigma-Aldrich (Saint-Quentin-Fallavier, FR).
Glacial acetic acid, Trizma® and Hexafluoroisopropanol (HFIP) were obtained from Sigma-Aldrich (FR). Deionized water (18 MΩ-cm) was obtained by using a Millipore Milli-Q Biocel A10 purification unit. Cuprisorb was purchased from Seachem. Ethanol (96.0%, EtOH), mEthanol (98.5%, MeOH) and acetonitrile (99.9%, ACN) were obtained from VWR international. NaCl (99%) was purchased from Alfa Aesar (FR). Azide monosaccharides (β-Dgalactopyranosyl azide, Gal-N 3 ; and β-D-glucopyranosyl azide (Glu-N 3 ) were obtained from Carbosynth (UK). Ammonium Acetate and Ammonium pyrrolidinedithiocarbamate, APDC, were purchased from Fisher Scientific (FR). RCA 120 and RCA 120 -Fluorescein were purchased from Eurobio (FR). Human serum from human male AB plasma, USA origin, sterile-filtered, was used as received from Sigma-Aldrich (FR). N,N,N′,N′′,N′′-pentamethyldiethylenetriamine (PMDETA) was purchased from Sigma-Aldrich (FR). CuSO 4 •5H 2 O was obtained from VWR (FR). Sodium ascorbate was obtained from Fisher Scientific (FR).
Glycidyl propargyl ether was obtained from Sigma-Aldrich (Saint-Quentin-Fallavier, FR).
Glacial acetic acid, Trizma® and Hexafluoroisopropanol (HFIP) were obtained from Sigma-Aldrich (FR). Deionized water (18 MΩ-cm) was obtained by using a Millipore Milli-Q Biocel A10 purification unit. Cuprisorb was purchased from Seachem. Ethanol (96.0%, EtOH), mEthanol (98.5%, MeOH) and acetonitrile (99.9%, ACN) were obtained from VWR international. NaCl (99%) was purchased from Alfa Aesar (FR). Azide monosaccharides (β-Dgalactopyranosyl azide, Gal-N 3 ; and β-D-glucopyranosyl azide (Glu-N 3 ) were obtained from Carbosynth (UK). Ammonium Acetate and Ammonium pyrrolidinedithiocarbamate, APDC, were purchased from Fisher Scientific (FR). RCA 120 and RCA 120 -Fluorescein were purchased from Eurobio (FR). Human serum from human male AB plasma, USA origin, sterile-filtered, was used as received from Sigma-Aldrich (FR). N,N,N′,N′′,N′′-pentamethyldiethylenetriamine (PMDETA) was purchased from Sigma-Aldrich (FR). CuSO 4 •5H 2 O was obtained from VWR (FR). Sodium ascorbate was obtained from Fisher Scientific (FR).
3
Recombinant Protein Expression in E. coli
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LB medium was purchased from Sigma-Aldrich (FR). Bacto-tryptone, and yeast extract were purchased from Biokar Diagnostics (FR). Ampicillin was obtained from Eurobio (FR). Glycerol and isopropyl β-D-thiogalactopyranoside (IPTG) were purchased from Euromedex (FR).
Complete mini EDTA-free protease inhibitors were purchased from Roche Diagnostics (D).
Bis(trifluoromethane)sulfonamide lithium salt was purchased from TCI Europe (BEL). Sodium
Complete mini EDTA-free protease inhibitors were purchased from Roche Diagnostics (D).
Bis(trifluoromethane)sulfonamide lithium salt was purchased from TCI Europe (BEL). Sodium
4
Reagents for Bacterial Culture Experiments
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Materials. LB medium was purchased from Sigma-Aldrich (Saint-Quentin-Fallavier, FR). Bacto-tryptone, and yeast extract were purchased from Biokar Diagnostics (Allone, FR). Ampicillin was obtained from Eurobio (Courtaboeuf, FR). Glycerol and isopropyl β-D-thiogalactopyranoside (IPTG) were purchased from Euromedex (Souffelweyersheim, FR). Complete mini EDTA-free protease inhibitors were purchased from Roche Diagnostics (Mannheim, D). Hydrogen peroxide and formic acid were purchased from Sigma-Aldrich (St. Louis, MO, USA). Glacial acetic acid was obtained from Fisher Scientific (USA). Deionized water (18 MΩ•cm) was obtained by passing in-house deionized water through a Millipore Milli-Q Biocel A10 purification unit.
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