Luria bertani lb

Manufactured by Merck Group

Sourced in United States, Italy

Luria Bertani (LB) is a commonly used growth medium for cultivating bacteria. It consists of a complex mixture of nutrients that support the growth of a wide range of bacterial species. The core function of LB is to provide the necessary components for bacterial growth, including carbon sources, nitrogen sources, vitamins, and minerals.

Automatically generated - may contain errors

Lab products found in correlation

33 protocols using luria bertani lb

Bacterial and Cell Culture Protocols

Check if the same lab product or an alternative is used in the 5 most similar protocols

EPEC E2348/69 strains (S1A Table) were grown in Luria Bertani (LB) (Sigma-Aldrich, St. Louis, Missouri, United States of America) broth or agar. Overnight bacterial cultures were grown at 37°C 180 revolutions per minute (rpm) shaking (liquid) or static (agar) and primed in Dulbecco’s Modified Eagle Medium (DMEM) as described below for infections.
SNU-C5, HT-29, SNU-C2B, COLO-320-HSR, and THP-1 cell lines were cultured in Roswell Park Memorial Institute (RPMI) medium (Sigma-Aldrich) with 10% (v/v) foetal bovine serum (FBS) (Gibco, Carlsbad, California, USA), 2 mM Glutamax (Gibco), 1 mM sodium pyruvate (Sigma-Aldrich), 10 mM N-2-hydroxyethylpiperazine-N-2-ethane sulphonic acid (HEPES) (Sigma-Aldrich), and 2,500 μg/ml glucose (Sigma-Aldrich). THP-1 cells were differentiated using 100 ng/ml phorbol 12-myristate 13-acetate (PMA) (Sigma-Aldrich) for 48 h and PMA withdrawn for 24 h. RPE cell line was cultured in DMEM/F-12 medium (Sigma-Aldrich) with 10% (v/v) FBS, 2 mM Glutamax, and 0.26% sodium bicarbonate (Gibco). HEK293E cell line was cultured in DMEM (high glucose, 4,500 mg/L) (Sigma-Aldrich) with 10% (v/v) FBS, 1 mM sodium pyruvate, and 10 mM HEPES (pH 7.4).

Zhong Q., Roumeliotis T.I., Kozik Z., Cepeda-Molero M., Fernández L.Á., Shenoy A.R., Bakal C., Frankel G, & Choudhary J.S. (2020). Clustering of Tir during enteropathogenic E. coli infection triggers calcium influx–dependent pyroptosis in intestinal epithelial cells. PLoS Biology, 18(12), e3000986.

+ Open protocol

+ Expand

Culturing C. violaceum for Bioreporter Assays

Check if the same lab product or an alternative is used in the 5 most similar protocols

In this study, a bioreporter strain of C. violaceum ATCC12472 was used. The strain was obtained from ATCC and stored at −80 °C supplemented with 20% glycerol, in the Department of Clinical Microbiology and Infectious Diseases, University of the Witwatersrand, Johannesburg, South Africa. Before experiments, the C. violaceum ATCC12472 was revived by culturing on Luria–Bertani (LB; Sigma Aldrich Co., Saint Louis, MO, USA) agar plates at 30 °C for 24 h.

Kamli M.R., Malik M.A., Srivastava V., Sabir J.S., Mattar E.H, & Ahmad A. (2021). Biogenic ZnO Nanoparticles Synthesized from Origanum vulgare Abrogates Quorum Sensing and Biofilm Formation in Opportunistic Pathogen Chromobacterium violaceum. Pharmaceutics, 13(11), 1743.

+ Open protocol

+ Expand

SARS-CoV-2 Pseudovirus Neutralization Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

Phosphate-buffered saline (PBS), lysozyme, Luria Bertani (LB), and HisTrap Ni-NTA columns are purchased from Sigma-Aldrich (St. Louis, MO). In-fusion cloning kit with Stellar cells was obtained from Takara Bio (San Jose, CA). E.coli (BL21-DE3 strains) was obtained from New England Biolabs Inc. (Ipswich, MA). Protease Inhibitor Tablets were obtained from Roche (Indianapolis, IN) and NHS-Cy5 was purchased from Lumiprobe Corp (Cockeysville, MD). Kanamycin Sulfate, B-PER Bacterial Protein Extraction Reagent, Penicillin-Streptomycin, Trypsin-EDTA, Dulbecco’s Modified Eagle Medium (DMEM, with high glucose), Puromycin Dihydrochloride, 4% paraformaldehyde in PBS, and Hoechst 33342 dye were purchased from Thermo Fisher Scientific (Waltham, MA). Fetal bovine serum was purchased from Hyclone (Logan, UT). Pseudovirus particles (PV) of SARS-CoV-2 (both Wuhan-Hu-1 and Delta variant B.1.617.2 strains) and 293T-hsACE2 T-hsACE2 cells were purchased from Integral Molecular (Philadelphia, PA). Free LCB1 mini-protein was custom-made and sold by Genscript Biotech Corp (Piscataway, NJ).

Chattaraj R., Kim C.Y., Lee D, & Hammer D.A. (2022). Recombinant Protein Micelles to Block Transduction by SARS-CoV-2 Pseudovirus. ACS nano, 16(10), 17466-17477.

+ Open protocol

+ Expand

Bacterial Strains and Culture Conditions

Check if the same lab product or an alternative is used in the 5 most similar protocols

STm wild type strain SL1344, aflagellate strain SL1344 (fliC::cat fljB::aph), SPI-1 mutant SL1344 SPI-1::aph and SPI-2 mutant SL1344 ssaV::aph were used together with B. subtilis NCTC 3610, EP E. coli O127:H6 E2348/69, K. pneumoniae NCTC 9633, P. aeruginosa wild type strain PA14 and aflagellate strain PA14 fliC::Tn Gm15^{54 (link)} and S. aureus NCTC 8532. Bacteria were cultured in Luria Bertani (LB; Sigma-Aldrich) broth at 37 °C with aeration, harvested at mid-log phase (OD₆₀₀ 0.5), washed three times in phosphate buffered saline and resuspended in 10% FBS/DMEM prior to inoculating epithelial cells. Bacteria were heat killed at 100 °C for 20 min and the absence of viable cells confirmed by overnight culture on LB plates.

Benedikz E.K., Bailey D., Cook C.N., Gonçalves-Carneiro D., Buckner M.M., Blair J.M., Wells T.J., Fletcher N.F., Goodall M., Flores-Langarica A., Kingsley R.A., Madsen J., Teeling J., Johnston S.L., MacLennan C.A., Balfe P., Henderson I.R., Piddock L.J., Cunningham A.F, & McKeating J.A. (2019). Bacterial flagellin promotes viral entry via an NF-kB and Toll Like Receptor 5 dependent pathway. Scientific Reports, 9, 7903.

+ Open protocol

+ Expand

Cultivation and Antibiotic Selection of Bacterial Strains

Check if the same lab product or an alternative is used in the 5 most similar protocols

S. sanguinis strain SK36 (kindly provided by Dr. Kilian) [27] and its derivatives were routinely cultured in Todd-Hewitt broth (TH, Becton Dickinson, NJ, USA) at 37°C. For a deoxyribonuclease (DNase) assay using agar plates, S. sanguinis strains as well as S. oralis NCTC 11427T/SK23 [27] , S. mutans MT8148 [28] (link), S. salivarius HHT [29] (link), S. parasanguinis ATCC 903 [27] , and S. sobrinus MT10186 [30] (link) were cultured in Brain Heart Infusion (BHI) broth (Becton Dickinson). The Escherichia coli strain TOP10 (Life Technologies, CA, USA) served as a host for derivatives of pSET6s and pAT18 [31] (link), [32] (link). The E. coli strain XL10-gold (Stratagene, CA, USA) was utilized as a host for the pQE30 derivatives (Qiagen, Germany). E. coli strains were cultured in Luria-Bertani (LB, Sigma Aldrich, MO, USA) medium at 37°C with constant agitation. Lactococcus lactis NZ9000 (kindly provided by Dr. Poolman) and its derivatives were grown in M17 broth (Becton Dickinson) containing 0.5% glucose (M17G, Wako, Japan) at 28°C. To select mutant strains, antibiotics were added to the media at the following concentrations: ampicillin (Wako); 100 µg/ml for E. coli, chloramphenicol (Sigma Aldrich); 10 µg/ml for E. coli and 5 µg/ml for S. sanguinis, and erythromycin (Sigma Aldrich); 150 µg/ml for E. coli and 1 µg/ml for L. lactis.

Morita C., Sumioka R., Nakata M., Okahashi N., Wada S., Yamashiro T., Hayashi M., Hamada S., Sumitomo T, & Kawabata S. (2014). Cell Wall-Anchored Nuclease of Streptococcus sanguinis Contributes to Escape from Neutrophil Extracellular Trap-Mediated Bacteriocidal Activity. PLoS ONE, 9(8), e103125.

+ Open protocol

+ Expand

Bacterial Infection of Intestinal Organoids

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

The bacterial strains used in this study are listed Table 2. Bacteria were routinely grown in Luria–Bertani (LB, Sigma Aldrich, Saint-Louis, MO, USA) broth with shaking at 150 rpm at 37 °C overnight. The day of infection, 2.10⁸ bacteria were added per well of 24-well plate for 90 min at 37 °C in a humidified atmosphere at 5% CO₂. After 90 min, infected organoids were washed with warm DMEM/F12 and then re-incubated further 90 min with fresh DMEM/F12. The time zero of post-infection (pi) was set at the beginning of the infection.Table 2

Bacterial strains and plasmids used in this study.

Strains	Relevant characteristic(s)	Source or reference
Strains MC1061	E. coli hsdR mcrB araD139 Δ(araABC-leu)7679 ΔlacX74 galU galK rpsL thi	[46 (link)]
STm	S. enterica subsp. enterica ser. Typhimurium 14028 wild-type strain	ATCC

Lacroix-Lamandé S., Bernardi O., Pezier T., Barilleau E., Burlaud-Gaillard J., Gagneux A., Velge P, & Wiedemann A. (2023). Differential Salmonella Typhimurium intracellular replication and host cell responses in caecal and ileal organoids derived from chicken. Veterinary Research, 54, 63.

+ Open protocol

+ Expand

Isolation and Maintenance of Pseudomonas Strains

Check if the same lab product or an alternative is used in the 5 most similar protocols

Pseudomonas spp. strains ITEM 17295, ITEM 17298 and ITEM 17299 were originally isolated from high-moisture mozzarella cheese [14 (link),15 (link)] and maintained at −80 °C as pure stock cultures in Nutrient Broth (NB; Oxoid S.p.A., Rodano, Milan, Italy) supplemented with glycerol 30% (v/v). The strains were routinely refreshed (30 °C, 24 h) by streaking onto Luria Bertani (LB; Sigma Aldrich, Milan, Italy) agar, and cultivated into 5 mL of LB broth for 16 h at 30 °C and 150 strokes to prepare the bacterial inocula for the subsequent experiments.

Quintieri L., Caputo L., De Angelis M, & Fanelli F. (2020). Genomic Analysis of Three Cheese-Borne Pseudomonas lactis with Biofilm and Spoilage-Associated Behavior. Microorganisms, 8(8), 1208.

+ Open protocol

+ Expand

HEK293 Cell Culture and Plasmid Transformation

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human embryonic kidney cell line 293 (HEK293) was kindly provided by Dr. Joseph Ruiz at Enzerna Biosciences (Raleigh, NC). Media and reagents for growing and maintaining cells were purchased from Life Technologies Corporation (Carlsbad, CA). The cells were maintained in Dulbecco’s Modified Eagle’s medium (DMEM) supplemented with 10% (by volume) fetal bovine serum and 1% penicillin-streptomycin. Cells were sustained in a humidified incubator at 37ºC and 5% CO₂. A 0.25% trypsin-EDTA solution was used for detachment of cells. One Shot Stbl3 Chemically Competent cells of Escherichia coli (Life Technologies Corporation) were used for constructing the mutagenesis plasmid. Bacteria carrying the plasmids were maintained in Luria Bertani (LB; Sigma-Aldrich, St. Louis, MO) agar or broth, and sustained in a humidified incubator at 37ºC.

Bandara A.B., Drake J.C., James C.C., Smyth J.W, & Brown D.A. (2021). Complex I protein NDUFS2 is vital for growth, ROS generation, membrane integrity, apoptosis, and mitochondrial energetics. Mitochondrion, 58, 160-168.

+ Open protocol

+ Expand

Bacterial Strain Cultivation Protocols

Check if the same lab product or an alternative is used in the 5 most similar protocols

For a list of all strains and plasmids
see Table S2.
E. coli and S. enterica cells were grown
at 37 °C in Luria-Bertani (LB, Sigma-Aldrich, USA) agar plates,
supplemented, when needed, with kanamycin (50 μg/mL, Roth) or
streptomycin (50 μg/mL, AppliChem). E. coli strains were fluorescently labeled with plasmid pGFP, S. enterica strains were fluorescently labeled with
plasmid pM965. A. Vinelandii cells
were grown on standard B media⁴² at 28
°C, the cells were tagged with GFP using the bjGFP plasmid, media
were supplemented with tetracycline (10 μg/mL, Sigma-Aldrich,
USA) if needed. V. natriegens was grown
on a LB3 medium (LB + NaCl fc. 3% w/v) at 37 °C and fluorescently
labeled using a sfGFP plasmid, and the expression was induced over
night with anhydrotetracycline (500 ng/mL, Adipogen. USA). For experiments,
all strains were grown to stationary phase via incubation for 48 h,
except E. coli strain ΔaroC ΔAAimporters, which was incubated
for 20 h at 28 °C.

Gäbelein C.G., Reiter M.A., Ernst C., Giger G.H, & Vorholt J.A. (2022). Engineering Endosymbiotic Growth of E. coli in Mammalian Cells. ACS Synthetic Biology, 11(10), 3388-3396.

+ Open protocol

+ Expand

Characterization of NorA Efflux Pump Mutants

Check if the same lab product or an alternative is used in the 5 most similar protocols

The S. aureus strains used in this study were an NCTC 8325-4 parental strain, a K1902 mutant (ΔnorA), and a K2378 mutant that overexpressed norA (norA++) from a multicopy plasmid (Felicetti et al., 2018 (link)). This strain was produced by cloning norA and its promoter into plasmid pCU1 and then introducing the construct into SA-K1902 (Augustin et al., 1992 (link)). The bacterial strains were donated by Dr. Glenn Kaatz of the Division of Infectious Diseases, School of Medicine, Wayne State University, United States, and were stored at the Laboratorio de Microbiología Básica y Aplicada, Universidad de Santiago de Chile. Bacterial cultures were grown in Mueller Hinton (MH) broth (Sigma Aldrich, United States) and Luria-Bertani (LB) (Sigma-Aldrich, United States) medium at 37°C with agitation (220 rpm). S. aureus K1902 and S. aureus K2378 cultures were supplemented with 10 μg/mL chloramphenicol.

Espinoza J., Urzúa A., Sanhueza L., Walter M., Fincheira P., Muñoz P., Mendoza L, & Wilkens M. (2019). Essential Oil, Extracts, and Sesquiterpenes Obtained From the Heartwood of Pilgerodendron uviferum Act as Potential Inhibitors of the Staphylococcus aureus NorA Multidrug Efflux Pump. Frontiers in Microbiology, 10, 337.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!