Structured Review

Becton Dickinson middlebrook 7h9 medium
Cell surface of Msmeg-PE11 is highly hydrophobic. ( a ) Pellicle formation of Msmeg-pVV or Msmeg-PE11 was monitored by growing standing cultures of the strains without shaking in <t>Middlebrook</t> <t>7H9</t> medium in absence of Tween 80 at 37 °C for various time points. ( b ) Biofilm formation was quantified by crystal violet staining for which Msmeg-pVV or Msmeg-PE11 cells were washed, stained with 1% crystal violet and ethanol extract was spectrophotometrically measured at 570 nm. ( c ) Msmeg-pVV or Msmeg-PE11 were cultured in 7H9 medium with (upper panel) or without (lower panel) 0.05% Tween-80 at 37 °C with shaking for 24 h and 48 h. Cultures were then allowed to settle at room temperature for 30 min. ( d ) In another experiment, Msmeg-pVV or Msmeg-PE11 were cultured in 7H9 broth with congo red (100 μg/ml) and 0.05% Tween 80 for 24 h, 48 h and 72 h at 37 °C. Cells were next washed and re-suspended in acetone. Congo red in the supernatant was spectrophotometrically measured at 490 nm. Data are representative of mean ± SD of three different experiments.
Middlebrook 7h9 Medium, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 97/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "PE11, a PE/PPE family protein of Mycobacterium tuberculosis is involved in cell wall remodeling and virulence"

Article Title: PE11, a PE/PPE family protein of Mycobacterium tuberculosis is involved in cell wall remodeling and virulence

Journal: Scientific Reports

doi: 10.1038/srep21624

Cell surface of Msmeg-PE11 is highly hydrophobic. ( a ) Pellicle formation of Msmeg-pVV or Msmeg-PE11 was monitored by growing standing cultures of the strains without shaking in Middlebrook 7H9 medium in absence of Tween 80 at 37 °C for various time points. ( b ) Biofilm formation was quantified by crystal violet staining for which Msmeg-pVV or Msmeg-PE11 cells were washed, stained with 1% crystal violet and ethanol extract was spectrophotometrically measured at 570 nm. ( c ) Msmeg-pVV or Msmeg-PE11 were cultured in 7H9 medium with (upper panel) or without (lower panel) 0.05% Tween-80 at 37 °C with shaking for 24 h and 48 h. Cultures were then allowed to settle at room temperature for 30 min. ( d ) In another experiment, Msmeg-pVV or Msmeg-PE11 were cultured in 7H9 broth with congo red (100 μg/ml) and 0.05% Tween 80 for 24 h, 48 h and 72 h at 37 °C. Cells were next washed and re-suspended in acetone. Congo red in the supernatant was spectrophotometrically measured at 490 nm. Data are representative of mean ± SD of three different experiments.
Figure Legend Snippet: Cell surface of Msmeg-PE11 is highly hydrophobic. ( a ) Pellicle formation of Msmeg-pVV or Msmeg-PE11 was monitored by growing standing cultures of the strains without shaking in Middlebrook 7H9 medium in absence of Tween 80 at 37 °C for various time points. ( b ) Biofilm formation was quantified by crystal violet staining for which Msmeg-pVV or Msmeg-PE11 cells were washed, stained with 1% crystal violet and ethanol extract was spectrophotometrically measured at 570 nm. ( c ) Msmeg-pVV or Msmeg-PE11 were cultured in 7H9 medium with (upper panel) or without (lower panel) 0.05% Tween-80 at 37 °C with shaking for 24 h and 48 h. Cultures were then allowed to settle at room temperature for 30 min. ( d ) In another experiment, Msmeg-pVV or Msmeg-PE11 were cultured in 7H9 broth with congo red (100 μg/ml) and 0.05% Tween 80 for 24 h, 48 h and 72 h at 37 °C. Cells were next washed and re-suspended in acetone. Congo red in the supernatant was spectrophotometrically measured at 490 nm. Data are representative of mean ± SD of three different experiments.

Techniques Used: Staining, Cell Culture

Related Articles

Electroporation:

Article Title: The 19 kDa Mycobacterium tuberculosis Lipoprotein (LpqH) Induces Macrophage Apoptosis through Extrinsic and Intrinsic Pathways: A Role for the Mitochondrial Apoptosis-Inducing Factor
Article Snippet: Mycobacteria and Isolation of LpqH A native Mycobacterium smegmatis strain (mc2155) and its counterpart transformed by electroporation with the plasmid p16R1 containing a 1.8 kb SmaI fragment that includes the structural gene encoding the LpqH lipoprotein were kindly donated by Y. Zhang (MRC Tuberculosis and Related Infections Unit, Hammersmith Hospital, London, UK). .. M. smegmatis strains were grown for 4-5 days in Middlebrook 7H9 medium (BBL, Becton-Dickinson, Cockeysville, MD, USA) supplemented with 2% glucose and hygromycin B (50 mg/mL).

Centrifugation:

Article Title: Variability in the virulence of specific Mycobacteriumtuberculosis clinical isolates alters the capacity of human dendritic cells to signal for T cells
Article Snippet: At day 7, MDDCs were collected by centrifugation and used for functional analyses. .. All strains were grown in Middlebrook 7H9 medium (BD-Diagnostic Systems, USA) supplemented with 10% Middlebrook Oleic Albumin Dextrose Catalase Growth Supplement (OADC) (BD-Diagnostic Systems, USA).

Article Title: The 19 kDa Mycobacterium tuberculosis Lipoprotein (LpqH) Induces Macrophage Apoptosis through Extrinsic and Intrinsic Pathways: A Role for the Mitochondrial Apoptosis-Inducing Factor
Article Snippet: M. smegmatis strains were grown for 4-5 days in Middlebrook 7H9 medium (BBL, Becton-Dickinson, Cockeysville, MD, USA) supplemented with 2% glucose and hygromycin B (50 mg/mL). .. Bacteria were harvested by centrifugation and washed with ice-cold sodium phosphate buffer (10 mM/L).

Selection:

Article Title: Variability in the virulence of specific Mycobacteriumtuberculosis clinical isolates alters the capacity of human dendritic cells to signal for T cells
Article Snippet: Alternatively, mononuclear cells isolated from healthy volunteers with positive tuberculin tests (range of 15 to 25 mm) were used to generate MDDCs and to purify CD4+ T cells by a magnetic bead approach using the CD4+ negative selection kit (Miltenyi, USA) according to manufacturer’s instructions. .. All strains were grown in Middlebrook 7H9 medium (BD-Diagnostic Systems, USA) supplemented with 10% Middlebrook Oleic Albumin Dextrose Catalase Growth Supplement (OADC) (BD-Diagnostic Systems, USA).

Sonication:

Article Title: Stable Extracellular RNA Fragments of Mycobacterium tuberculosis Induce Early Apoptosis in Human Monocytes via a Caspase-8 Dependent Mechanism
Article Snippet: Growth of M. tuberculosis To prepare bacilli for monocyte infections, M. tuberculosis H37Rv (kindly provided by Laboratorio de Micobacterias, Instituto Nacional de Salud, Bogota, Colombia) was grown as a pellicle in Middlebrook 7H9 medium (Becton Dickinson, Sparks, MD) supplemented with glycerol (Promega, Madison, WI), oleic acid-albumin-dextrose-catalase (OADC) (Becton Dickinson), and Tween 80 (Becton Dickinson). .. To disrupt bacterial clumps, M. tuberculosis was suspended in RPMI-1640 with 20% glycerol and probe sonicated (Model CV33 Sonics Vibra Cell, Newtown, CT) five times at 2.5 W output for 1 min at 4°C.

Article Title: The 19 kDa Mycobacterium tuberculosis Lipoprotein (LpqH) Induces Macrophage Apoptosis through Extrinsic and Intrinsic Pathways: A Role for the Mitochondrial Apoptosis-Inducing Factor
Article Snippet: M. smegmatis strains were grown for 4-5 days in Middlebrook 7H9 medium (BBL, Becton-Dickinson, Cockeysville, MD, USA) supplemented with 2% glucose and hygromycin B (50 mg/mL). .. To isolate LpqH, cell-wall fractions were obtained by sonication of bacteria at 20 KHz in iced water (five cycles of 5 min each).

Isolation:

Article Title: Pyrazinoic Acid Inhibits Mycobacterial Coenzyme A Biosynthesis by Binding to Aspartate Decarboxylase PanD
Article Snippet: M. bovis BCG (ATCC 35734) and M. tuberculosis H37Rv (ATCC 27294) strains were maintained in complete Middlebrook 7H9 medium (BD Difco) supplemented with 0.05% (v/v) Tween 80 (Sigma), 0.5% (v/v) glycerol (Fisher Scientific), and 10% (v/v) Middlebrook albumin-dextrosecatalase (BD Difco) at 37 °C with agitation at 80 rpm. .. POA-resistant strains used for this study, M. bovis BCG POA2.2 and M. bovis BCG POA1.3, were isolated and described in ref .

Article Title: Variability in the virulence of specific Mycobacteriumtuberculosis clinical isolates alters the capacity of human dendritic cells to signal for T cells
Article Snippet: Alternatively, mononuclear cells isolated from healthy volunteers with positive tuberculin tests (range of 15 to 25 mm) were used to generate MDDCs and to purify CD4+ T cells by a magnetic bead approach using the CD4+ negative selection kit (Miltenyi, USA) according to manufacturer’s instructions. .. All strains were grown in Middlebrook 7H9 medium (BD-Diagnostic Systems, USA) supplemented with 10% Middlebrook Oleic Albumin Dextrose Catalase Growth Supplement (OADC) (BD-Diagnostic Systems, USA).

Article Title: The 19 kDa Mycobacterium tuberculosis Lipoprotein (LpqH) Induces Macrophage Apoptosis through Extrinsic and Intrinsic Pathways: A Role for the Mitochondrial Apoptosis-Inducing Factor
Article Snippet: Paragraph title: 2.2. Mycobacteria and Isolation of LpqH ... M. smegmatis strains were grown for 4-5 days in Middlebrook 7H9 medium (BBL, Becton-Dickinson, Cockeysville, MD, USA) supplemented with 2% glucose and hygromycin B (50 mg/mL).

Article Title: The Phenolic Glycolipid of Mycobacterium tuberculosis Differentially Modulates the Early Host Cytokine Response but Does Not in Itself Confer Hypervirulence
Article Snippet: Paragraph title: Isolation of RNA. ... Bacterial cultures were grown in triplicate in Middlebrook 7H9 medium (Becton Dickinson, Sparks, MD) supplemented with 10% oleic acid-albumin-dextrose-catalase (BD Biosciences, Sparks, MD), 0.2% glycerol, and 0.05% Tween 80 to late exponential phase.

Functional Assay:

Article Title: Variability in the virulence of specific Mycobacteriumtuberculosis clinical isolates alters the capacity of human dendritic cells to signal for T cells
Article Snippet: At day 7, MDDCs were collected by centrifugation and used for functional analyses. .. All strains were grown in Middlebrook 7H9 medium (BD-Diagnostic Systems, USA) supplemented with 10% Middlebrook Oleic Albumin Dextrose Catalase Growth Supplement (OADC) (BD-Diagnostic Systems, USA).

Cell Culture:

Article Title: Membrane Type 1 Matrix Metalloproteinase Regulates Monocyte Migration and Collagen Destruction in Tuberculosis
Article Snippet: .. M. tuberculosis H37Rv was cultured in Middlebrook 7H9 medium (BD Biosciences, Oxford, U.K.). .. M. tuberculosis in mid-log growth at an OD of 0.60 (Biowave cell density meter, WPA) was used for infecting monocytes immediately after adhesion purification at a multiplicity of infection (MOI) of 1.

Mouse Assay:

Article Title: Variability in the virulence of specific Mycobacteriumtuberculosis clinical isolates alters the capacity of human dendritic cells to signal for T cells
Article Snippet: The Mtb strain H37Rv was used in order to establish an experimental reference for the comparative analyses done with the clinical isolate strains Ph1 (highly virulent, with no induction of protective immune response in mice) and Ph4 (less virulent, provoking a protective adaptive immune response). .. All strains were grown in Middlebrook 7H9 medium (BD-Diagnostic Systems, USA) supplemented with 10% Middlebrook Oleic Albumin Dextrose Catalase Growth Supplement (OADC) (BD-Diagnostic Systems, USA).

Sequencing:

Article Title: Mycobacterial Cell Wall Synthesis Inhibitors Cause Lethal ATP Burst
Article Snippet: M. bovis BCG Pasteur (ATCC #35734) p iniBAC -RFP harbors the coding sequence of the red fluorescent protein mCherry under control of the iniBAC promoter and was constructed as previously described ( ). .. Liquid cultures were grown in complete Middlebrook 7H9 medium (BD Difco) supplemented with 0.05% Tween 80, 0.4% glycerol, and 10% albumin-dextrose-catalase enrichment (Becton Dickinson) at 37°C and 80 rpm.

Concentration Assay:

Article Title: Pyrazinoic Acid Inhibits Mycobacterial Coenzyme A Biosynthesis by Binding to Aspartate Decarboxylase PanD
Article Snippet: M. bovis BCG (ATCC 35734) and M. tuberculosis H37Rv (ATCC 27294) strains were maintained in complete Middlebrook 7H9 medium (BD Difco) supplemented with 0.05% (v/v) Tween 80 (Sigma), 0.5% (v/v) glycerol (Fisher Scientific), and 10% (v/v) Middlebrook albumin-dextrosecatalase (BD Difco) at 37 °C with agitation at 80 rpm. .. Pyrazinoic acid was freshly dissolved in 90% DMSO at a concentration of 0.5 M and sterilized using 0.2 μ m PTFE membrane filters (Acrodisc PALL).

Article Title: Noncytotoxic Pyrrolobenzodiazepine–Ciprofloxacin Conjugate with Activity against Mycobacterium tuberculosis
Article Snippet: Reagents used were Middlebrook 7H9 medium (BD Difco), OADC supplemented with 0.5% glycerol (24388.295, VWR Chemicals) and 0.2% Tween 80 (P1754, Sigma), CAMR Mycobacterium medium MOD2 (CMM MOD2), ciprofloxacin (Sigma-Aldrich, cat no. 33434), and 0.02% resazurin solution (10 mg resazurin sodium salt (R7017, Sigma-Aldrich) in 50 mL phosphate-buffered saline, pH 7.4 (Severn Biotech) containing 5% Tween 80). .. All of the antibiotics were dissolved in dimethyl sulphoxide (DMSO) (Sigma-Aldrich D2650) to a stock concentration of 10 mg/mL.

Incubation:

Article Title: The Phenolic Glycolipid of Mycobacterium tuberculosis Differentially Modulates the Early Host Cytokine Response but Does Not in Itself Confer Hypervirulence
Article Snippet: Bacterial cultures were grown in triplicate in Middlebrook 7H9 medium (Becton Dickinson, Sparks, MD) supplemented with 10% oleic acid-albumin-dextrose-catalase (BD Biosciences, Sparks, MD), 0.2% glycerol, and 0.05% Tween 80 to late exponential phase. .. After a 5-min incubation on ice, the supernatant was transferred to a 1.5-ml microcentrifuge tube and centrifuged at 14,000 rpm for 15 min at 4°C, followed by transfer of the supernatant to a new 1.5-ml tube.

other:

Article Title: A bacterial hemerythrin-like protein MsmHr inhibits the SigF-dependent hydrogen peroxide response in mycobacteria
Article Snippet: Culture medium and growth conditions M. smegmatis cultures were grown in Middlebrook 7H9 medium (Becton Dickinson, Sparks, MD) supplemented with ADS enrichment (Albumin-Dextrose Saline containing 5% (w/v) Bovine serum albumin fraction V, 2% (w/v) D-Dextrose and 8.1% (w/v) NaCl) (Jacobs et al., ), 0.05% (v/v) Tween 80, and 0.5% (v/v) glycerol (Beijing Modern Eastern Finechemical Co. Ltd, Beijing).

Article Title: In Vitro Activities of Enantiopure and Racemic 1′-Acetoxychavicol Acetate against Clinical Isolates of Mycobacterium tuberculosis
Article Snippet: Bacterial Strains and Culture Condition M. tuberculosis H37Ra ATCC 25177 and H37Rv ATCC 27294 including clinical isolates were grown in Middlebrook 7H9 medium (Becton Dickinson, Sparks, MD, USA) supplemented with 10% Oleic Acid-Albumin-Dextrose-Catalase (OADC) and 0.05% Tween-80 and on solid agar medium Middlebrook 7H10 medium (Becton Dickinson) supplemented with 10% OADC.

Infection:

Article Title: Variability in the virulence of specific Mycobacteriumtuberculosis clinical isolates alters the capacity of human dendritic cells to signal for T cells
Article Snippet: All strains were grown in Middlebrook 7H9 medium (BD-Diagnostic Systems, USA) supplemented with 10% Middlebrook Oleic Albumin Dextrose Catalase Growth Supplement (OADC) (BD-Diagnostic Systems, USA). .. Internalisation of Mtb and infection of MDDCs - Bacterial aliquots (H37Rv, Ph1, and Ph4) were thawed at room temperature, and the bacterial aggregate declumping was achieved by vortexing with borosilicate beads for 5 min and centrifuging at 2040 × g for 5 min. MDDC infection was performed at a multiplicity of infection (MOI) of 5 (5 bacteria to 1 cell).

Article Title: Membrane Type 1 Matrix Metalloproteinase Regulates Monocyte Migration and Collagen Destruction in Tuberculosis
Article Snippet: M. tuberculosis H37Rv was cultured in Middlebrook 7H9 medium (BD Biosciences, Oxford, U.K.). .. M. tuberculosis in mid-log growth at an OD of 0.60 (Biowave cell density meter, WPA) was used for infecting monocytes immediately after adhesion purification at a multiplicity of infection (MOI) of 1.

Polyacrylamide Gel Electrophoresis:

Article Title: The 19 kDa Mycobacterium tuberculosis Lipoprotein (LpqH) Induces Macrophage Apoptosis through Extrinsic and Intrinsic Pathways: A Role for the Mitochondrial Apoptosis-Inducing Factor
Article Snippet: M. smegmatis strains were grown for 4-5 days in Middlebrook 7H9 medium (BBL, Becton-Dickinson, Cockeysville, MD, USA) supplemented with 2% glucose and hygromycin B (50 mg/mL). .. Proteins were resolved by 12% or 15% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to nitrocellulose.

Colony-forming Unit Assay:

Article Title: Variability in the virulence of specific Mycobacteriumtuberculosis clinical isolates alters the capacity of human dendritic cells to signal for T cells
Article Snippet: All strains were grown in Middlebrook 7H9 medium (BD-Diagnostic Systems, USA) supplemented with 10% Middlebrook Oleic Albumin Dextrose Catalase Growth Supplement (OADC) (BD-Diagnostic Systems, USA). .. Bacillary viability was tested using the colony forming unit (CFU) assay, growing serial dilutions in 7H10 agar plates (BD-Diagnostic Systems, USA) supplemented with 10% OADC (BD-Diagnostic Systems, USA) for 14 and 21 days.

Modification:

Article Title: Noncytotoxic Pyrrolobenzodiazepine–Ciprofloxacin Conjugate with Activity against Mycobacterium tuberculosis
Article Snippet: Bacterial Strains and Reagents Mycobacteriumtuberculosis strains, H37Rv, 1192/015, and 08/00483E (multidrug-resistant (MDR)), were used in the modified resazurin microtiter assay (REMA) plate method. .. Reagents used were Middlebrook 7H9 medium (BD Difco), OADC supplemented with 0.5% glycerol (24388.295, VWR Chemicals) and 0.2% Tween 80 (P1754, Sigma), CAMR Mycobacterium medium MOD2 (CMM MOD2), ciprofloxacin (Sigma-Aldrich, cat no. 33434), and 0.02% resazurin solution (10 mg resazurin sodium salt (R7017, Sigma-Aldrich) in 50 mL phosphate-buffered saline, pH 7.4 (Severn Biotech) containing 5% Tween 80).

Construct:

Article Title: Mycobacterial Cell Wall Synthesis Inhibitors Cause Lethal ATP Burst
Article Snippet: M. bovis BCG Pasteur (ATCC #35734) p iniBAC -RFP harbors the coding sequence of the red fluorescent protein mCherry under control of the iniBAC promoter and was constructed as previously described ( ). .. Liquid cultures were grown in complete Middlebrook 7H9 medium (BD Difco) supplemented with 0.05% Tween 80, 0.4% glycerol, and 10% albumin-dextrose-catalase enrichment (Becton Dickinson) at 37°C and 80 rpm.

Article Title: Differential Effects of Mycobacterium bovis BCG on Macrophages and Dendritic Cells from Murine Spleen
Article Snippet: Bacterial Strains and Culture Conditions M.bovis BCG Pasteur 1173P2 and rBCG-GFP were kindly provided by Dr. Xiaoming Zhang (Institut Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai). rBCG::Ag85A was constructed by our laboratory. .. M.bovis BCG Pasteur 1173P2, rBCG-GFP and rBCG::Ag85A were grown with gentle agitation (80 rpm) in Middlebrook 7H9 medium (BD, Sparks, MD, USA) supplemented with 0.05% Tween 80 and 10% acid-albumin-dextrose-catalase complex (ADC), or on solid Middlebrook 7H10 medium (BD, Sparks, MD, USA) supplemented with 0.05% Tween 80 and 10% oleic acid-albumin-dextrose-catalase complex (OADC).

Transformation Assay:

Article Title: The 19 kDa Mycobacterium tuberculosis Lipoprotein (LpqH) Induces Macrophage Apoptosis through Extrinsic and Intrinsic Pathways: A Role for the Mitochondrial Apoptosis-Inducing Factor
Article Snippet: Mycobacteria and Isolation of LpqH A native Mycobacterium smegmatis strain (mc2155) and its counterpart transformed by electroporation with the plasmid p16R1 containing a 1.8 kb SmaI fragment that includes the structural gene encoding the LpqH lipoprotein were kindly donated by Y. Zhang (MRC Tuberculosis and Related Infections Unit, Hammersmith Hospital, London, UK). .. M. smegmatis strains were grown for 4-5 days in Middlebrook 7H9 medium (BBL, Becton-Dickinson, Cockeysville, MD, USA) supplemented with 2% glucose and hygromycin B (50 mg/mL).

Article Title: PE11, a PE/PPE family protein of Mycobacterium tuberculosis is involved in cell wall remodeling and virulence
Article Snippet: .. M. smegmatis culture and transformation M. smegmatis mc2 155 bacteria were grown in Middlebrook 7H9 medium (BD Difco, USA) supplemented with 10% OADC (HiMedia, India), 0.5% glycerol, and 0.05% Tween 80 (Fisher Scientific, USA). ..

Purification:

Article Title: Membrane Type 1 Matrix Metalloproteinase Regulates Monocyte Migration and Collagen Destruction in Tuberculosis
Article Snippet: M. tuberculosis H37Rv was cultured in Middlebrook 7H9 medium (BD Biosciences, Oxford, U.K.). .. M. tuberculosis in mid-log growth at an OD of 0.60 (Biowave cell density meter, WPA) was used for infecting monocytes immediately after adhesion purification at a multiplicity of infection (MOI) of 1.

SDS Page:

Article Title: The 19 kDa Mycobacterium tuberculosis Lipoprotein (LpqH) Induces Macrophage Apoptosis through Extrinsic and Intrinsic Pathways: A Role for the Mitochondrial Apoptosis-Inducing Factor
Article Snippet: M. smegmatis strains were grown for 4-5 days in Middlebrook 7H9 medium (BBL, Becton-Dickinson, Cockeysville, MD, USA) supplemented with 2% glucose and hygromycin B (50 mg/mL). .. Proteins were resolved by 12% or 15% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to nitrocellulose.

Plasmid Preparation:

Article Title: The 19 kDa Mycobacterium tuberculosis Lipoprotein (LpqH) Induces Macrophage Apoptosis through Extrinsic and Intrinsic Pathways: A Role for the Mitochondrial Apoptosis-Inducing Factor
Article Snippet: Mycobacteria and Isolation of LpqH A native Mycobacterium smegmatis strain (mc2155) and its counterpart transformed by electroporation with the plasmid p16R1 containing a 1.8 kb SmaI fragment that includes the structural gene encoding the LpqH lipoprotein were kindly donated by Y. Zhang (MRC Tuberculosis and Related Infections Unit, Hammersmith Hospital, London, UK). .. M. smegmatis strains were grown for 4-5 days in Middlebrook 7H9 medium (BBL, Becton-Dickinson, Cockeysville, MD, USA) supplemented with 2% glucose and hygromycin B (50 mg/mL).

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  • 93
    Becton Dickinson 7h9 liquid medium
    Growth curves of the WT, ΔMycP 3 mutant and the two complementation strains, ΔMycP3 ms ::pr1MycP3 ms and ΔMycP3 ms ::pr2MycP3 ms under Fe-free <t>7H9</t> ( a ), Fe-free Sauton’s media ( b ). The growth curves were done in triplicate, error bars show standard deviation
    7h9 Liquid Medium, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/7h9 liquid medium/product/Becton Dickinson
    Average 93 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    7h9 liquid medium - by Bioz Stars, 2020-04
    93/100 stars
      Buy from Supplier

    97
    Becton Dickinson middlebrook 7h9 medium
    Cell surface of Msmeg-PE11 is highly hydrophobic. ( a ) Pellicle formation of Msmeg-pVV or Msmeg-PE11 was monitored by growing standing cultures of the strains without shaking in <t>Middlebrook</t> <t>7H9</t> medium in absence of Tween 80 at 37 °C for various time points. ( b ) Biofilm formation was quantified by crystal violet staining for which Msmeg-pVV or Msmeg-PE11 cells were washed, stained with 1% crystal violet and ethanol extract was spectrophotometrically measured at 570 nm. ( c ) Msmeg-pVV or Msmeg-PE11 were cultured in 7H9 medium with (upper panel) or without (lower panel) 0.05% Tween-80 at 37 °C with shaking for 24 h and 48 h. Cultures were then allowed to settle at room temperature for 30 min. ( d ) In another experiment, Msmeg-pVV or Msmeg-PE11 were cultured in 7H9 broth with congo red (100 μg/ml) and 0.05% Tween 80 for 24 h, 48 h and 72 h at 37 °C. Cells were next washed and re-suspended in acetone. Congo red in the supernatant was spectrophotometrically measured at 490 nm. Data are representative of mean ± SD of three different experiments.
    Middlebrook 7h9 Medium, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 97/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/middlebrook 7h9 medium/product/Becton Dickinson
    Average 97 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    middlebrook 7h9 medium - by Bioz Stars, 2020-04
    97/100 stars
      Buy from Supplier

    Image Search Results


    Growth curves of the WT, ΔMycP 3 mutant and the two complementation strains, ΔMycP3 ms ::pr1MycP3 ms and ΔMycP3 ms ::pr2MycP3 ms under Fe-free 7H9 ( a ), Fe-free Sauton’s media ( b ). The growth curves were done in triplicate, error bars show standard deviation

    Journal: BMC Research Notes

    Article Title: Two promoters in the esx-3 gene cluster of Mycobacterium smegmatis respond inversely to different iron concentrations in vitro

    doi: 10.1186/s13104-017-2752-0

    Figure Lengend Snippet: Growth curves of the WT, ΔMycP 3 mutant and the two complementation strains, ΔMycP3 ms ::pr1MycP3 ms and ΔMycP3 ms ::pr2MycP3 ms under Fe-free 7H9 ( a ), Fe-free Sauton’s media ( b ). The growth curves were done in triplicate, error bars show standard deviation

    Article Snippet: M. smegmatis was cultured using both Middlebrook 7H9 liquid medium (Becton–Dickinson, USA, Catalogue No. 221832) and Difco 7H11 solid medium (Becton–Dickinson, USA, Catalogue No. 212304) both supplemented with 0.05% (v/v) Tween 80 (Sigma Aldrich, USA, Catalogue No. P1754), 0.5% (w/v) glucose (Sigma Aldrich, USA, Catalogue No. 47829), and 0.5% (v/v) glycerol (Sigma-Aldrich, USA, Catalogue No. G5516).

    Techniques: Mutagenesis, Mass Spectrometry, Standard Deviation

    The comparison of the intracellular iron levels in WT ms , ΔMycP3 ms , ΔMycP3 ms ::pr1MycP3 ms and ΔMycP3 ms ::pr2MycP3 ms strains under 7H9, Fe-depleted 7H9 and Fe rescued 7H9 media. The error bars show standard error of the mean (n = 4). The p values obtained using two-way ANOVA statistical analysis between different culturing conditions for all four strains are smaller than 0.0001 ( *** ), an example is shown for the WT ms

    Journal: BMC Research Notes

    Article Title: Two promoters in the esx-3 gene cluster of Mycobacterium smegmatis respond inversely to different iron concentrations in vitro

    doi: 10.1186/s13104-017-2752-0

    Figure Lengend Snippet: The comparison of the intracellular iron levels in WT ms , ΔMycP3 ms , ΔMycP3 ms ::pr1MycP3 ms and ΔMycP3 ms ::pr2MycP3 ms strains under 7H9, Fe-depleted 7H9 and Fe rescued 7H9 media. The error bars show standard error of the mean (n = 4). The p values obtained using two-way ANOVA statistical analysis between different culturing conditions for all four strains are smaller than 0.0001 ( *** ), an example is shown for the WT ms

    Article Snippet: M. smegmatis was cultured using both Middlebrook 7H9 liquid medium (Becton–Dickinson, USA, Catalogue No. 221832) and Difco 7H11 solid medium (Becton–Dickinson, USA, Catalogue No. 212304) both supplemented with 0.05% (v/v) Tween 80 (Sigma Aldrich, USA, Catalogue No. P1754), 0.5% (w/v) glucose (Sigma Aldrich, USA, Catalogue No. 47829), and 0.5% (v/v) glycerol (Sigma-Aldrich, USA, Catalogue No. G5516).

    Techniques: Mass Spectrometry

    Growth curves of the WT, ΔMycP 3 mutant and the two complementation strains, ΔMycP3 ms ::pr1MycP3 ms and ΔMycP3 ms ::pr2MycP3 ms under Fe-depleted 7H9 ( a ), and Fe-depleted Sauton’s media ( b ). The growth curves were done in triplicate, error bars show standard deviation

    Journal: BMC Research Notes

    Article Title: Two promoters in the esx-3 gene cluster of Mycobacterium smegmatis respond inversely to different iron concentrations in vitro

    doi: 10.1186/s13104-017-2752-0

    Figure Lengend Snippet: Growth curves of the WT, ΔMycP 3 mutant and the two complementation strains, ΔMycP3 ms ::pr1MycP3 ms and ΔMycP3 ms ::pr2MycP3 ms under Fe-depleted 7H9 ( a ), and Fe-depleted Sauton’s media ( b ). The growth curves were done in triplicate, error bars show standard deviation

    Article Snippet: M. smegmatis was cultured using both Middlebrook 7H9 liquid medium (Becton–Dickinson, USA, Catalogue No. 221832) and Difco 7H11 solid medium (Becton–Dickinson, USA, Catalogue No. 212304) both supplemented with 0.05% (v/v) Tween 80 (Sigma Aldrich, USA, Catalogue No. P1754), 0.5% (w/v) glucose (Sigma Aldrich, USA, Catalogue No. 47829), and 0.5% (v/v) glycerol (Sigma-Aldrich, USA, Catalogue No. G5516).

    Techniques: Mutagenesis, Mass Spectrometry, Standard Deviation

    Gene expression analysis of mycP 3 in WT ms , ΔMycP 3ms , ΔMycP 3ms ::Pr1MycP 3 , and ΔMycP 3ms ::Pr2MycP 3 strains under normal 7H9 (iron rich), Fe-free 7H9, and Fe-rescued 7H9 media. The results were normalized against the RNA copy number of sigA . The p values obtained using two-way ANOVA statistical analysis (n = 3) (*p

    Journal: BMC Research Notes

    Article Title: Two promoters in the esx-3 gene cluster of Mycobacterium smegmatis respond inversely to different iron concentrations in vitro

    doi: 10.1186/s13104-017-2752-0

    Figure Lengend Snippet: Gene expression analysis of mycP 3 in WT ms , ΔMycP 3ms , ΔMycP 3ms ::Pr1MycP 3 , and ΔMycP 3ms ::Pr2MycP 3 strains under normal 7H9 (iron rich), Fe-free 7H9, and Fe-rescued 7H9 media. The results were normalized against the RNA copy number of sigA . The p values obtained using two-way ANOVA statistical analysis (n = 3) (*p

    Article Snippet: M. smegmatis was cultured using both Middlebrook 7H9 liquid medium (Becton–Dickinson, USA, Catalogue No. 221832) and Difco 7H11 solid medium (Becton–Dickinson, USA, Catalogue No. 212304) both supplemented with 0.05% (v/v) Tween 80 (Sigma Aldrich, USA, Catalogue No. P1754), 0.5% (w/v) glucose (Sigma Aldrich, USA, Catalogue No. 47829), and 0.5% (v/v) glycerol (Sigma-Aldrich, USA, Catalogue No. G5516).

    Techniques: Expressing, Mass Spectrometry

    Mutation of fadA5 does not cause cholesterol toxicity. Strains were grown to exponential phase in 7H9 medium supplemented with glycerol and dextrose. Cultures were diluted into 7H9 medium containing glycerol and dextrose with or without cholesterol (1 mg ml −1 in 20% Tween). Values are for the wild-type H37Rv strain without cholesterol, wild-type H37Rv with cholesterol, the fadA5 mutant without cholesterol, and fadA5 mutant with cholesterol, as indicated. Data represent the results of the experiment done in triplicate.

    Journal: Infection and Immunity

    Article Title: A Thiolase of Mycobacterium tuberculosis Is Required for Virulence and Production of Androstenedione and Androstadienedione from Cholesterol ▿ Is Required for Virulence and Production of Androstenedione and Androstadienedione from Cholesterol ▿ †

    doi: 10.1128/IAI.00893-09

    Figure Lengend Snippet: Mutation of fadA5 does not cause cholesterol toxicity. Strains were grown to exponential phase in 7H9 medium supplemented with glycerol and dextrose. Cultures were diluted into 7H9 medium containing glycerol and dextrose with or without cholesterol (1 mg ml −1 in 20% Tween). Values are for the wild-type H37Rv strain without cholesterol, wild-type H37Rv with cholesterol, the fadA5 mutant without cholesterol, and fadA5 mutant with cholesterol, as indicated. Data represent the results of the experiment done in triplicate.

    Article Snippet: M. tuberculosis cultures were grown at 37°C in Middlebrook 7H9 liquid medium (Becton Dickinson), supplemented with 0.05% Tween-80, 10% albumin-dextrose-NaCl complex (ADN) , and 0.2% glycerol, or on Middlebrook 7H10 plates supplemented the same way.

    Techniques: Mutagenesis

    fadA5 is required for growth on cholesterol as the sole carbon source. The strains were grown in 7H9 medium containing 1 mg ml −1 (2.6 mM) cholesterol in tyloxapol or 7H9 medium containing only tyloxapol at 37°C. The wild-type H37Rv (Rv), fadA5 mutant, and complemented fadA5 (fadA5comp) strains were grown with and without cholesterol as indicated on the graph. Data represent results of each experiment run in duplicate.

    Journal: Infection and Immunity

    Article Title: A Thiolase of Mycobacterium tuberculosis Is Required for Virulence and Production of Androstenedione and Androstadienedione from Cholesterol ▿ Is Required for Virulence and Production of Androstenedione and Androstadienedione from Cholesterol ▿ †

    doi: 10.1128/IAI.00893-09

    Figure Lengend Snippet: fadA5 is required for growth on cholesterol as the sole carbon source. The strains were grown in 7H9 medium containing 1 mg ml −1 (2.6 mM) cholesterol in tyloxapol or 7H9 medium containing only tyloxapol at 37°C. The wild-type H37Rv (Rv), fadA5 mutant, and complemented fadA5 (fadA5comp) strains were grown with and without cholesterol as indicated on the graph. Data represent results of each experiment run in duplicate.

    Article Snippet: M. tuberculosis cultures were grown at 37°C in Middlebrook 7H9 liquid medium (Becton Dickinson), supplemented with 0.05% Tween-80, 10% albumin-dextrose-NaCl complex (ADN) , and 0.2% glycerol, or on Middlebrook 7H10 plates supplemented the same way.

    Techniques: Mutagenesis

    Cell surface of Msmeg-PE11 is highly hydrophobic. ( a ) Pellicle formation of Msmeg-pVV or Msmeg-PE11 was monitored by growing standing cultures of the strains without shaking in Middlebrook 7H9 medium in absence of Tween 80 at 37 °C for various time points. ( b ) Biofilm formation was quantified by crystal violet staining for which Msmeg-pVV or Msmeg-PE11 cells were washed, stained with 1% crystal violet and ethanol extract was spectrophotometrically measured at 570 nm. ( c ) Msmeg-pVV or Msmeg-PE11 were cultured in 7H9 medium with (upper panel) or without (lower panel) 0.05% Tween-80 at 37 °C with shaking for 24 h and 48 h. Cultures were then allowed to settle at room temperature for 30 min. ( d ) In another experiment, Msmeg-pVV or Msmeg-PE11 were cultured in 7H9 broth with congo red (100 μg/ml) and 0.05% Tween 80 for 24 h, 48 h and 72 h at 37 °C. Cells were next washed and re-suspended in acetone. Congo red in the supernatant was spectrophotometrically measured at 490 nm. Data are representative of mean ± SD of three different experiments.

    Journal: Scientific Reports

    Article Title: PE11, a PE/PPE family protein of Mycobacterium tuberculosis is involved in cell wall remodeling and virulence

    doi: 10.1038/srep21624

    Figure Lengend Snippet: Cell surface of Msmeg-PE11 is highly hydrophobic. ( a ) Pellicle formation of Msmeg-pVV or Msmeg-PE11 was monitored by growing standing cultures of the strains without shaking in Middlebrook 7H9 medium in absence of Tween 80 at 37 °C for various time points. ( b ) Biofilm formation was quantified by crystal violet staining for which Msmeg-pVV or Msmeg-PE11 cells were washed, stained with 1% crystal violet and ethanol extract was spectrophotometrically measured at 570 nm. ( c ) Msmeg-pVV or Msmeg-PE11 were cultured in 7H9 medium with (upper panel) or without (lower panel) 0.05% Tween-80 at 37 °C with shaking for 24 h and 48 h. Cultures were then allowed to settle at room temperature for 30 min. ( d ) In another experiment, Msmeg-pVV or Msmeg-PE11 were cultured in 7H9 broth with congo red (100 μg/ml) and 0.05% Tween 80 for 24 h, 48 h and 72 h at 37 °C. Cells were next washed and re-suspended in acetone. Congo red in the supernatant was spectrophotometrically measured at 490 nm. Data are representative of mean ± SD of three different experiments.

    Article Snippet: M. smegmatis culture and transformation M. smegmatis mc2 155 bacteria were grown in Middlebrook 7H9 medium (BD Difco, USA) supplemented with 10% OADC (HiMedia, India), 0.5% glycerol, and 0.05% Tween 80 (Fisher Scientific, USA).

    Techniques: Staining, Cell Culture