Lb broth

Manufactured by BD

Sourced in United States

LB broth is a commonly used nutrient-rich medium for the growth and cultivation of various microorganisms, including bacteria. It provides the essential nutrients and growth factors required for the proliferation of microbial cultures in a laboratory setting.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

Luria-Bertani broth (LB; (14 citations) Difco LB broth (11 citations) LB broth Miller (8 citations) Lennox LB broth (5 citations) Difco lysogeny broth (LB; (4 citations) Luria broth (LB; (3 citations) Luria-Bertani (Miller) Broth (LB) (3 citations) Luria-Bertani broth (LB) medium (2 citations) Difco LB broth Miller (Luria−Bertani), (2 citations) LB broth powder (2 citations)

98 protocols using lb broth

Competitive E. coli Infection in Mice

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

The E. coli strains were routinely grown aerobically at 37°C in LB broth (BD Biosciences) or on LB plates. When necessary, antibiotics were added to the medium at the following concentrations: 0.1 mg/ml carbenicillin, 0.05 mg/ml kanamycin, 0.03 mg/ml chloramphenicol, or 0.1 mg/ml streptomycin. When grown for competitive infection, strains were grown in a hypoxia chamber (0.8% oxygen) at 37°C in LB broth (BD Biosciences).
(ii) Animal experiments. All experiments in this study were approved by the Institutional Animal Care and Use Committee at the University of California at Davis. Female C57BL/6J wild-type mice, aged 8 to 10 weeks, were obtained from The Jackson Laboratory.

Cevallos S.A., Lee J.Y., Tiffany C.R., Byndloss A.J., Johnston L., Byndloss M.X, & Bäumler A.J. (2019). Increased Epithelial Oxygenation Links Colitis to an Expansion of Tumorigenic Bacteria. mBio, 10(5), e02244-19.

+ Open protocol

+ Expand

Bacterial Strain Construction and Media

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

Detailed bacterial strain information, including growth media and temperature, is provided in Table S1. Where listed, growth media were prepared according manufacturer recommendations: LB Broth and LB Broth supplemented with 0.3 mM thymine (BD Biosciences 244610, Alfa Aesar A15879), Brain Heart Infusion (BD Biosciences 237500), Tryptic Soy Broth (BD Biosciences 211825), Nutrient Broth (BD Biosciences 234000), and Gutnick Minimal Media (1.0 g/L K₂SO₄, 13.5 g/L K₂HPO₄, 4.7 g/L KH₂PO₄, 0.1 g/L of MgSO₄-7H₂O, 10 mM NH₄Cl as a nitrogen source, and 0.4% w/v glucose as a carbon source) (Gutnick et al., 1969 (link)), Terrific Broth (BD Biosciences 243820).
E. coli lptD4213 ΔthyA was constructed by moving ΔthyA::Km^r (Bell-Pedersen et al., 1991 (link)) into E. coli lptD4213 by P1 transduction.

Martin JK I.I., Sheehan J.P., Bratton B.P., Moore G.M., Mateus A., Li S.H., Kim H., Rabinowitz J.D., Typas A., Savitski M.M., Wilson M.Z, & Gitai Z. (2020). A Dual-Mechanism Antibiotic Kills Gram-Negative Bacteria and Avoids Drug Resistance. Cell, 181(7), 1518-1532.e14.

+ Open protocol

+ Expand

Protein Extraction from Hypermutator Strains

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

Sample preparation and protein extraction were carried out according to previously described methods (Han et al., 2013 (link)), with some modifications. The four hypermutator strains and Salmonella Typhimurium LT2 were each streaked on LB agar (BD) plates and incubated at 37°C. Afterward, a single colony from each strain was subcultured in 5 ml of LB broth (BD) at 37°C for 14 h. Then, 0.05 ml of the subcultures were added into 10 ml of LB broth (BD) and incubated at 37°C for another 5 h with shaking at 120 rpm. Cells were harvested by centrifugation at 4°C for 5 min at 15,000 × g, and the pellets were washed three times using 4°C distilled water.
The cells were then resuspended in lysis buffer containing 7 M urea, 2% (w/v) chaps, 2% (v/v, pH 3–10) pharmalyte, 1% (w/v) dithiothreitol, and 5 mM pefabloc (proteinase inhibitor). Subsequently, the cell suspensions were incubated on ice for 15 min and sonicated until clear solutions were obtained. After centrifugation (4°C, 15,000 × g, 15 min), the supernatants were collected. Protein concentrations in the supernatants were measured using a Bradford Protein Assay Kit (TaKaRa).

Sheng H., Huang J., Han Z., Liu M., Lü Z., Zhang Q., Zhang J., Yang J., Cui S, & Yang B. (2020). Genes and Proteomes Associated With Increased Mutation Frequency and Multidrug Resistance of Naturally Occurring Mismatch Repair-Deficient Salmonella Hypermutators. Frontiers in Microbiology, 11, 770.

+ Open protocol

+ Expand

E. coli Nissle 1917 Mutant Characterization

Cited 1 time

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

E. coli Nissle 1917 is a commensal human isolate (43 (link)) that has been marketed as a probiotic. The E. coli Nissle 1917 cydA napA narG narZ mutant and the E. coli Nissle 1917 cydA mutant used in this study have been described previously (29 (link)). E. coli strains were routinely grown aerobically at 37°C in LB broth (BD Biosciences) or on LB agar plates. When appropriate, antibiotics were added to the medium at the following concentrations: 0.1 mg/ml carbenicillin and 0.05 mg/ml kanamycin. For competitive infection experiments, strains were grown in a hypoxia chamber (0.8% oxygen) at 37°C in LB broth (BD Biosciences).

Cevallos S.A., Lee J.Y., Velazquez E.M., Foegeding N.J., Shelton C.D., Tiffany C.R., Parry B.H., Stull-Lane A.R., Olsan E.E., Savage H.P., Nguyen H., Ghanaat S.S., Byndloss A.J., Agu I.O., Tsolis R.M., Byndloss M.X, & Bäumler A.J. (2021). 5-Aminosalicylic Acid Ameliorates Colitis and Checks Dysbiotic Escherichia coli Expansion by Activating PPAR-γ Signaling in the Intestinal Epithelium. mBio, 12(1), e03227-20.

+ Open protocol

+ Expand

Downshock Experiments for Bacterial Survival

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

Downshock experiments were conducted as previously described [2 (link), 30 (link), 31 (link)]. A single colony was used to inoculate an overnight culture in Luria-Bertani Broth (LB Broth, BD Biosciences, San Jose, CA) supplemented with ampicillin (100µg/mL), the overnight culture was subsequently used to inoculate (1:20) a culture in LB Broth with 250 mM NaCl and ampicillin. The resulting culture was grown to an OD₆₀₀ of approximately 0.5 and induced with 0.1 mM isopropyl-beta-D-thiogalactopyranoside (IPTG) for 30 min. Following 30 min of induction, the culture was diluted (1:40) into 1:1 LB Broth and deionized water or isotonically into LB Broth with 250 mM NaCl, and allowed to recover for 30 m in a shaking incubator. After hypoosmotic downshock or isotonic dilution, bacterial cultures were serially diluted and plated on LB plates supplemented with ampicillin (100µg/mL). Plates containing between 25 and 250 colonies were used to determine the colony forming units (CFU) per millimeter of media. Percent recovery was defined as the CFU of the downshocked culture divided by the CFU of the isotonic dilution. Six trials for each mutation were conducted.

Malcolm H.R., Blount P, & Maurer J.A. (2014). The mechanosensitive channel of small conductance (MscS) functions as a Jack-In-The Box. Biochimica et biophysica acta, 1848(1 0 0), 159-166.

+ Open protocol

+ Expand

Cultivation and Maintenance of Bacterial Strains

Cited 1 time

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

H37Rv (H37Rv), M. bovis BCG, E.coli DH5α and P. aeruginosa PAO1 were gifts from Professor Vojo Deretic,^{37 (link),38 (link)}M. tuberculosis katGS315T (katG-S315T) was a gift from Professor Alex Pym ^{36 (link)}. S. aureus USA300 LAC was a gift from Professor Pamela Hall ^{39 (link)}. All bacterial cultures were grown at 37°C with shaking. Mycobacterium cultures were prepared by thawing frozen stock aliquots: H37Rv and katG-S315T were grown in 7H9 Middlebrook liquid medium supplemented with oleic acid, albumin, dextrose and catalase (Becton Dickinson, Inc., Sparks, MD), 0.5% glycerol and 0.05% Tween 80. BCG was grown in the same culture medium omitting oleic acid. Escherichia coli DH5α was grown overnight in LB broth (Becton Dickinson), Pseudomonas aeruginosa strain PAO1 was grown overnight in LB broth supplemented with 1.76% NaCl and 1% glycerol, and Staphylococcus aureus USA300 LAC was grown overnight in BBL Trypticase soybroth (Becton Dickinson).

Choi S.W., Maiga M., Maiga M.C., Atudorei V., Sharp Z.D., Bishai W.R, & Timmins G.S. (2014). Rapid in vivo detection of isoniazid-sensitive Mycobacterium tuberculosis by breath test. Nature communications, 5, 4989.

+ Open protocol

+ Expand

Quantitative RT-PCR Analysis of bla_OXA-55-like Transcription in S. algae

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

Quantitative reverse transcription-PCR (RT-PCR) was performed for S. algae clade isolates to compare bla_OXA-55-like transcription levels. The isolates were grown in LB broth (Becton Dickinson and Co.) for 24 h at 37°C with shaking at 160 rpm and harvested at an optical density at 600 nm (OD₆₀₀) of 1.0. The RNA was extracted using the RNeasy minikit (Qiagen, Hilden, Germany), then used to generate cDNA with PrimeScript RT master mix (TaKaRa Bio Inc.). Quantitative PCR was performed using SYBR green PCR master mix (Applied Biosystems, Foster City, CA) with the primer pair bla_OXA-55-like_Forward_Primer (5′-GTTGGTTGGAGTTGGACGAC-3′) and bla_OXA-55-like_Reverse_Primer (5′-TGCTTGAGCACCTGTTTCAC-3′) on the Applied Biosystems 7500 Fast system (Applied Biosystems). Amplification of the rpoB gene was simultaneously performed with the rpoB_Forward_Primer (5′-TTTGATCCCATTCCTTGAGC-3′) and rpoB_Reverse_Primer (5′-CCACCAGAGGCTTCTCTGAC-3′). The bla_OXA-55-like transcription levels of the isolates were compared using the threshold cycle (ΔΔC_T) method (21 (link)). The amplification efficiency of the quantitative PCR for rpoB and bla_OXA-55-like was verified with 10-fold serially diluted TUM17384 total RNA ranging from 10⁰ ng to 10⁻⁵ng per assay, which demonstrated that the ratio of amplification efficiency of bla_OXA-55-like to rpoB was 0.98.

Ohama Y., Aoki K., Harada S., Nagasawa T., Sawabe T., Nonaka L., Moriya K., Ishii Y, & Tateda K. (2021). Genetic Environment Surrounding blaOXA-55-like in Clinical Isolates of Shewanella algae Clade and Enhanced Expression of blaOXA-55-like in a Carbapenem-Resistant Isolate. mSphere, 6(5), e00593-21.

+ Open protocol

+ Expand

Salmonella enterica Typhimurium Growth Protocol

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

In this study, Salmonella enterica serovar Typhimurium (ATCC 14028) (ST) was used. ST was grown in Luria-Bertani (LB) (Becton, Dickinson and Co., Franklin Lakes, NJ, USA) agar at 37 °C under aerobic conditions (Thermo Fisher Scientific Inc., Marietta, OH, USA). All experiments described were performed in LB broth (Becton, Dickinson and Co., Franklin Lakes, NJ, USA) and were incubated under aerobic conditions for 24 h in 37 °C with continuous aeration at 150 rpm through the use of a shaking incubator.

Alvarado-Martinez Z., Bravo P., Kennedy N.F., Krishna M., Hussain S., Young A.C, & Biswas D. (2020). Antimicrobial and Antivirulence Impacts of Phenolics on Salmonella Enterica Serovar Typhimurium. Antibiotics, 9(10), 668.

+ Open protocol

+ Expand

Construction and Characterization of Stress-Resistant Bacterial Strains

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 in Table S3. Salmonella or Escherichia coli (E. coli) strains were grown in LB broth (Becton Dickinson; Franklin Lakes, NJ, USA) or agar plates at 37 °C. Antibiotics were supplied, if required (chloramphenicol, 34 μg/ml; kanamycin, 50 μg/ml; ampicillin, 100 μg/ml). KST0649 and KST0651 strains were constructed using the λ red recombination system as described previously^{54 (link)}. The KST0650 strain was isolated from its parent strain (KST0649) using RMT. In Brief, KST0649 was cultured in LB broth (OD₆₀₀ = 0.5), followed by irradiation using a Co-60 gamma irradiator (Advanced Research Technology Institute; JeonEupSi, Korea) for 0.5 h (1.2 kGy). Irradiated KST0649 was spread on LB agar with 1 mM H₂O₂, followed by incubation for 48 h at 37 °C. The colonies were randomly selected and cultured in LB broth for 24 h and the high H₂O₂-resistant strains were selected.

Gao S., Jung J.H., Lin S.M., Jang A.Y., Zhi Y., Bum Ahn K., Ji H.J., Hyang Lim J., Guo H., Choy H.E., Lim S, & Seo H.S. (2020). Development of Oxytolerant Salmonella typhimurium Using Radiation Mutation Technology (RMT) for Cancer Therapy. Scientific Reports, 10, 3764.

+ Open protocol

+ Expand

Bacterial Viability Assay with 2,4-DAPG

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

Bacterial cells were grown in LB-broth (Becton-Dickinson, Sparks, MD, USA) to mid-log phase, then centrifuged (9391× g for 10 min), re-suspended in HEPES buffer (10 mM) and adjusted to an optical density corresponding to 10⁸ CFU/mL. 2,4-DAPG was then mixed with E. coli and S. aureus 209P at concentrations corresponding to their MIC. After a 2-h incubation at 25 °C, the cells were washed with distilled water. For microscopy, the suspension was mixed with 0.4% agarose, LIVE/DEAD dyes, and plated. Fluorescence microscopy of stained bacteria was performed using a Zeiss Axio Imager M2 fluorescent microscope (Zeiss, Oberkochen, Germany) equipped with filter sets for simultaneous viewing of Syto 9 and PI fluorescence.

Julian W.T., Vasilchenko A.V., Shpindyuk D.D., Poshvina D.V, & Vasilchenko A.S. (2020). Bacterial-Derived Plant Protection Metabolite 2,4-Diacetylphloroglucinol: Effects on Bacterial Cells at Inhibitory and Subinhibitory Concentrations. Biomolecules, 11(1), 13.

+ 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!