The recombinant bacteria were cultured in Luria-Bertani (LB, Sangon Biotech Co., Ltd., Shanghai, China) liquid medium containing 100.0 µg/mL kanamycin, at 37 °C shaken (150 rpm) overnight. The cultured bacteria were transferred to fresh LB medium (10.0 mL, 100.0 µg/mL kanamycin), and cultured at 37 °C with shaking (150 rpm) until OD600 reached 0.6. Isopropyl β-D-1-thiogalactopyranoside (IPTG; Merck, Germany) was added to a total concentration of 0.5 mmol/L, and incubated at 26 °C with shaking (120 rpm) for 12–16 h. The induced recombinant bacteria were adjusted to OD600 1.0 and stored at 4 °C for further use.
Luria bertani lb
Manufactured by Sangon
Sourced in China
Luria-Bertani (LB) is a commonly used growth medium for culturing bacteria. It provides the necessary nutrients and growth factors to support the cultivation of a wide range of bacterial species. LB is a versatile and widely used product in microbiological and molecular biology laboratories.
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Lab products found in correlation
Isopropyl β d 1 thiogalactopyranoside (iptg), by Merck Group (1 mentions)
Salmonella typhimurium, by American Type Culture Collection (1 mentions)
Klebsiella pneumoniae, by American Type Culture Collection (1 mentions)
Listeria monocytogenes, by American Type Culture Collection (1 mentions)
Vibrio parahaemolyticus, by American Type Culture Collection (1 mentions)
Shewanella putrefaciens, by American Type Culture Collection (1 mentions)
Pseudomonas aeruginosa pao1, by American Type Culture Collection (1 mentions)
Enterobacter sakazakii, by American Type Culture Collection (1 mentions)
Pseudomonas fluorescens, by American Type Culture Collection (1 mentions)
Melatonin, by Merck Group (1 mentions)
4 protocols using luria bertani lb
1
Recombinant Bacteria Induction Protocol
2
Antimicrobial Spectrum Screening of Bacterial Strains
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Staphylococcus aureus (RN 4220) and Escherichia coli (ATCC 25922) were the primary indicator bacteria. Samomella enteritidis (CCTCC AB 94018), Klebsiella Pneumoniae (ATCC 10031), Enterobacter Sakazakii (ATCC 29544), Vibrio parahaemolyticus (ATCC 10031), Listeria monocytogenes (ATCC 19115), Pseudomonas aeruginosa PAO1, Salmonella typhimurium (ATCC 14028), Shewanella putrefaciens (ATCC 8071), and Pseudomonas fluorescens (ATCC 13525) were purchased from China Center of Industrial Culture Collection (CICC). Lactiplantibacillus plantarum DMDL 9010, Lacticaseibacillus rhamnosus B1107, Bacillus coagulans 13002, Bacillus amyloliquefaciens K1, and Bacillus licheniformis SG18 were isolated and preserved in Guangdong Microbial Culture Collection center (GDMCC). All strains were used as indicator bacteria for antimicrobial spectrum detection.
The medium of tryptone-glucose-yeast extract (TGYE) and nutritional broth (NB) was used for screening the antimicrobial strains from the sample; tryptic soy broth (TSB), brain heart infusion (BHI), Luria-Bertani (LB), and de Man, Rogosa and Sharpe (MRS) purchased from Sangon Biotech (Shanghai) were used as the cultures for different indicator bacteria.
The medium of tryptone-glucose-yeast extract (TGYE) and nutritional broth (NB) was used for screening the antimicrobial strains from the sample; tryptic soy broth (TSB), brain heart infusion (BHI), Luria-Bertani (LB), and de Man, Rogosa and Sharpe (MRS) purchased from Sangon Biotech (Shanghai) were used as the cultures for different indicator bacteria.
3
ETEC Infection of 3D4/21 Cells with Melatonin
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The ETEC F4ac C83902, ETEC F4ac ΔLT and ETEC F4ac ΔSTb were used in this study, among which wild strains were E. coli W25K (O149: K91, K88ac; LT, STb, EAST) (Ren et al., 2014a ), and the knockout strains were provided by Professor Guoqiang Zhu, College of Veterinary Medicine, Yangzhou University, China. The ETEC cells were cultured in 10 mL Luria–Bertani (LB) (Sangon Biotech, Shanghai, China) at 37 °C and harvested at the log phase. The 3D4/21 cells were infected with ETEC for 1 h (multiplicity of infection [MOI] = 10:1), and culture supernatant and cells were collected. In some experiments, 3D4/21 cells were pre-treated with 1 mM melatonin (Sigma–Aldrich, St Louis, USA) for 18 h followed by ETEC infection.
4
Heterologous Expression of Synthetic NGAL
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The synthetic NGAL gene (Generay Biotechnology, Shanghai, China) was 549 bp in the pET32a vector, and then was transformed into E. coli BL21 to express protein. The bacteria were grown in 5 mL Luria-Bertani (LB, Sangon Biotech, Shanghai, China) Broth medium supplemented with 100 mg mL -1 ampicillin at 37°C and shaking at 220 rpm overnight.
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