Staphylococcus aureus (ATCC 25923), which is sensitive to vancomycin (MIC: 0.9 μg/ml), but not sensitive to ceftriaxone, was used in this study [23 (link)]. S. aureus was cultured on trypticase soy agar (Oxoid item number LP0042), transferred to trypticase soy broth. The bacteriawere incubated for 12 h at 37 °C. After centrifuging at 2,000 g for 5 min, the pellets wereresuspended and diluted to different levels, the concentration of the bacteria was determined at 550 nm with a spectrophotometer (UV1600, Mapada Equipment Co. Ltd., Shanghai, China) and further estimated by plating on trypticase soy agar plates. Based on the previous report, 100 μl 1 × 107 CFU/mL of S. aureus ATCC 25923 was used to create a reliable infection rate [24 (link)].
Lp0042
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
The LP0042 is a laboratory equipment product from Thermo Fisher Scientific. It is a precision instrument designed for performing specific laboratory operations. The core function of the LP0042 is to enable accurate and reliable measurements or analyses within a controlled laboratory environment. No further details on the intended use or application of this product are provided.
Automatically generated - may contain errors
Lab products found in correlation
Lp0021, by Thermo Fisher Scientific (3 mentions)
Lp0011, by Thermo Fisher Scientific (2 mentions)
M6030, by Merck Group (1 mentions)
Lp0011b, by Thermo Fisher Scientific (1 mentions)
Tryptone, by Thermo Fisher Scientific (1 mentions)
Cm0131, by Thermo Fisher Scientific (1 mentions)
Lp0013, by Thermo Fisher Scientific (1 mentions)
Cm0129, by Thermo Fisher Scientific (1 mentions)
6 protocols using lp0042
1
Preparing Vancomycin-Sensitive S. aureus
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Staphylococcus aureus (ATCC 25923), which is sensitive to vancomycin (MIC: 0.9 μg/ml), but not sensitive to ceftriaxone, was used in this study [23 (link)]. S. aureus was cultured on trypticase soy agar (Oxoid item number LP0042), transferred to trypticase soy broth. The bacteriawere incubated for 12 h at 37 °C. After centrifuging at 2,000 g for 5 min, the pellets wereresuspended and diluted to different levels, the concentration of the bacteria was determined at 550 nm with a spectrophotometer (UV1600, Mapada Equipment Co. Ltd., Shanghai, China) and further estimated by plating on trypticase soy agar plates. Based on the previous report, 100 μl 1 × 107 CFU/mL of S. aureus ATCC 25923 was used to create a reliable infection rate [24 (link)].
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Staphylococcus aureus (ATCC 25923), which is sensitive to vancomycin (MIC: 0.9 μg/ml), but not sensitive to ceftriaxone, was used in this study [23 (link)]. S. aureus was cultured on trypticase soy agar (Oxoid item number LP0042), transferred to trypticase soy broth. The bacteriawere incubated for 12 h at 37 °C. After centrifuging at 2,000 g for 5 min, the pellets wereresuspended and diluted to different levels, the concentration of the bacteria was determined at 550 nm with a spectrophotometer (UV1600, Mapada Equipment Co. Ltd., Shanghai, China) and further estimated by plating on trypticase soy agar plates. Based on the previous report, 100 μl 1 × 107 CFU/mL of S. aureus ATCC 25923 was used to create a reliable infection rate [24 (link)].
2
Preparation of Luria-Bertani and M9 Media
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Luria-Bertani (LB) medium contained the following ingredients (per litre): 5 g yeast extract (L21; Oxoid), 10 g tryptone (LP0042; Oxoid), various amounts of NaCl as detailed in the growth sections below, and the pH was adjusted to 7.8 with 1 M NaOH before autoclaving. For solid medium, 15 g of bacteriological agar No1 (LP0011; Oxoid) was also added per litre. M9 Minimal Salts medium (M-6030; Sigma) was supplemented with 1 mM MgSO4, 1 g NH4Cl l−1, 0.1 mM CaCl2 and 3 g glucose l−1. The pH was adjusted to between 8 and 9 using 1 M NaOH. NaCl was added to the basal M9 medium to generate the range of salinities required, using the concentrations specified below. It was sterilized by autoclaving.
3
Bacteriophage Infection of E. coli K-12
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The bacterial strain used in this study was E. coli E. coli E. coli
Bacterial strains used in this study were grown in LB [10 g l−1 Tryptone (Oxoid: LP0042), 10 g l−1 NaCl (Oxoid: LP0005) and 5 g l−1 Yeast extract (Oxoid: LP0021)] as either liquid culture (with shaking at 100 r.p.m.) or solid culture [supplemented with 15 g l−1 agar (Oxoid: LP0011B)] at 37 °C. Phages were suspended in phage buffer (10 mM Tris.HCl pH 7.5, 8 mM MgSO4).
Bacterial strains used in this study were grown in LB [10 g l−1 Tryptone (Oxoid: LP0042), 10 g l−1 NaCl (Oxoid: LP0005) and 5 g l−1 Yeast extract (Oxoid: LP0021)] as either liquid culture (with shaking at 100 r.p.m.) or solid culture [supplemented with 15 g l−1 agar (Oxoid: LP0011B)] at 37 °C. Phages were suspended in phage buffer (10 mM Tris.HCl pH 7.5, 8 mM MgSO4).
4
Cultivation and Washing of L. varians GY32
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L. varians GY32 was isolated from heavy metals-polluted sediment and preserved in our laboratory17 (link). Strain GY32 was cultivated aerobically in Luria–Bertani broth (LB) for 12 h (30 °C, 120 rpm) in flasks. The LB (1 L) contained 5 g yeast extract (LP0021, Oxoid, United Kingdom), 10 g Tryptone (LP0042, Oxoid, United Kingdom), and 5 g NaCl. Before being used for inoculation to MFCs or further tests, the LB-cultivated bacteria cells were washed in sterilized phosphate buffered saline (PBS) or deionized water for at least three times by centrifugation at 8000×g for 5 min. PBS used here (pH 7.2, 1 L) contained 8 g NaCl (C111545, Aladdin, China), 0.2 g KCl (P112133, Aladdin, China), 3.63 g Na2HPO4·12H2O (S112623, Aladdin, China), 0.24 g KH2PO4 (P104071, Aladdin, China), the pH was adjusted by HCl (10011008, Hushi, China).
5
Characterizing Phage Interactions with Staphylococci
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Phage host range was determined by spotting serial dilutions on double-layer agar plates with tryptone soya agar (TSA) bottom agar (CM0131, Oxoid, Basingstoke, United Kingdom) and 0.6% (w/v) top Agar No. 1 (LP0011, Oxoid) with the addition of CaCl2 to a concentration of 2 mM. The growth properties and adsorption efficiency of phages were determined in tryptone soya broth (TSB) (CM0129, Oxoid), with 2 mM CaCl2 as described previously [17 (link)] with minor modification for staphylococci [18 (link)]. The ability to grow on different media was further tested on meat peptone agar (MPA) and double-concentrated yeast extract-tryptone (2× YT) agar composed of 1.6% (w/v) tryptone (LP0042, Oxoid), 1.0% (w/v) yeast extract (LP0021, Oxoid), 0.5% (w/v) NaCl, and 1.5% (w/v) agar (LP0013, Oxoid), pH 7.0. A one-step growth curve and phage burst size were determined by the procedure described previously [17 (link)]. Free plasmacoagulase and clumping factor tests were performed as described previously [19 (link),20 (link)].
6
Growth and Infection of E. coli with Bacteriophages
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The bacterial strain used in this study was Escherichia coli K-12 MG1655 (28) . The bacteriophage used in this study were T4 phage which is a Virulent Myovirus that infects E. coli (DSM4505) and T7 phage which is a Virulent Podovirus that infects E. coli (DSM4623). These phages were obtained from the Leibniz-Institut DSMZ -Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH.
Bacterial strains used in this study were grown in LB (10 g L -1 Tryptone (Oxoid: LP0042), 10 g L -1 NaCl (Oxoid: LP0005) and 5 g L -1 Yeast extract (Oxoid: LP0021)) as either liquid culture (with shaking at 100 rpm) or solid culture (supplemented with 15 g L -1 agar (Oxoid: LP0011B)) at 37 o C.
Phages were suspended in lambda diluent (10 mM Tris.HCl pH 7.5, 8 mM MgSO4).
Bacterial strains used in this study were grown in LB (10 g L -1 Tryptone (Oxoid: LP0042), 10 g L -1 NaCl (Oxoid: LP0005) and 5 g L -1 Yeast extract (Oxoid: LP0021)) as either liquid culture (with shaking at 100 rpm) or solid culture (supplemented with 15 g L -1 agar (Oxoid: LP0011B)) at 37 o C.
Phages were suspended in lambda diluent (10 mM Tris.HCl pH 7.5, 8 mM MgSO4).
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