Structured Review

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
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1) Product Images from "Two promoters in the esx-3 gene cluster of Mycobacterium smegmatis respond inversely to different iron concentrations in vitro"

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

Journal: BMC Research Notes

doi: 10.1186/s13104-017-2752-0

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
Figure Legend 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

Techniques Used: 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
Figure Legend 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

Techniques Used: 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
Figure Legend 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

Techniques Used: 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
Figure Legend 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

Techniques Used: Expressing, Mass Spectrometry

2) Product Images from "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 ▿ †"

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 ▿ †

Journal: Infection and Immunity

doi: 10.1128/IAI.00893-09

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.
Figure Legend 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.

Techniques Used: 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.
Figure Legend 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.

Techniques Used: Mutagenesis

3) Product Images from "Antimicrobial Treatment Improves Mycobacterial Survival in Nonpermissive Growth Conditions"

Article Title: Antimicrobial Treatment Improves Mycobacterial Survival in Nonpermissive Growth Conditions

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.02774-13

RaaS is important for persistence of M. tuberculosis in prolonged stationary phase and in human macrophages. (A) Wild-type, Δ raaS Mtb , and Δ raaS Mtb com strains were incubated in 7H9 medium without shaking at 37°C for 7 months. **, Δ raaS Mtb had a significant survival defect ( P
Figure Legend Snippet: RaaS is important for persistence of M. tuberculosis in prolonged stationary phase and in human macrophages. (A) Wild-type, Δ raaS Mtb , and Δ raaS Mtb com strains were incubated in 7H9 medium without shaking at 37°C for 7 months. **, Δ raaS Mtb had a significant survival defect ( P

Techniques Used: Incubation

4) Product Images from "The volatiles of pathogenic and nonpathogenic mycobacteria and related bacteria"

Article Title: The volatiles of pathogenic and nonpathogenic mycobacteria and related bacteria

Journal: Beilstein Journal of Organic Chemistry

doi: 10.3762/bjoc.8.31

Volatiles released by M. tuberculosis grown in a 7H9 broth liquid medium.
Figure Legend Snippet: Volatiles released by M. tuberculosis grown in a 7H9 broth liquid medium.

Techniques Used:

Related Articles

Clone Assay:

Article Title: MspA Nanopores from Subunit Dimers
Article Snippet: Bacterial Strains and Growth Conditions M. smegmatis ML16, which lacks the porin genes mspA , mspC , and mspD was grown at 37°C in 7H9 liquid medium (BD Biosciences) supplemented with 0.2% glycerol and 0.05% Tween 80 or on 7H10 agar (BD Biosciences) supplemented with 0.2% glycerol, unless otherwise indicated. .. Escherichia coli DH5α was used for cloning experiments and was routinely grown in Luria-Bertani broth (LB) at 37°C.

Centrifugation:

Article Title: Increasing Prevalence of Nontuberculous Mycobacteria in Respiratory Specimens from US-Affiliated Pacific Island Jurisdictions 1
Article Snippet: Laboratory Testing Specimens were digested by using the standard NALC-NaOH method, followed by centrifugation ( ). .. The remaining sediment was resuspended and used to inoculate 7H9 liquid medium (BACTEC MGIT 960; Becton Dickinson, Franklin Lakes, NJ, USA), broth medium, and solid medium (Middlebrook 7H10 agar slant; Becton Dickinson).

Incubation:

Article Title: Direct detection of Mycobacterium tuberculosis rifampin resistance in bio-safe stained sputum smears
Article Snippet: .. For culture, 45% of the suspension (135 μl) was inoculated into 7H9 liquid medium supplemented with Albumin Dextrose complex and PANTA (polymyxin B, amphotericin B, nalidixic acid, trimethoprim and azlocillin) supplement (Becton Dickinson, USA), and the tubes were incubated at 37°C for upto 8 weeks. .. The positivity in the 7H9 liquid culture was first identified on the basis of turbidity of the medium, which was then confirmed for presence of M . tuberculosis by ZN smear microscopy and IS6110 PCR (M . tuberculosis complex specific PCR) [ ].

Article Title: Increasing Prevalence of Nontuberculous Mycobacteria in Respiratory Specimens from US-Affiliated Pacific Island Jurisdictions 1
Article Snippet: The remaining sediment was resuspended and used to inoculate 7H9 liquid medium (BACTEC MGIT 960; Becton Dickinson, Franklin Lakes, NJ, USA), broth medium, and solid medium (Middlebrook 7H10 agar slant; Becton Dickinson). .. The broth medium was incubated in a MGIT 960 semiautomated system (Becton Dickinson) and the 7H10 agar slant was incubated at 35°C in an incubator containing 5% CO2 .

Article Title: The volatiles of pathogenic and nonpathogenic mycobacteria and related bacteria
Article Snippet: Bacterial colonies from a 7H11 solid medium (Becton, Dickinson & Co., Sparks, USA, see ) or bacteria culture (100 µL) from a 7H9 liquid medium (Becton, Dickinson & Co., Sparks, USA, see ) were inoculated on a 7H11 solid medium and were spread out with sterile disposable loops to cover the entire plate/medium surface. .. Plates were wrapped in parafilm and aerobically incubated at 28–37 °C.

Infection:

Article Title: Mycobacterium tuberculosis from chronic murine infections that grows in liquid but not on solid medium
Article Snippet: Culture media and bacteria The media used were 7H9 liquid medium with 10% albumin, dextrose, catalase supplement and 0.05% Tween 80, and 7H11 agar medium with 10% oleic acid, albumin, dextrose, catalase supplement (Becton Dickinson, Oxford, UK). .. The two experiments both used 6-week, female Balbc mice, which were infected, with a mouse passaged H37Rv strain of M. tuberculosis suspended in 0.1% gelatin.

Article Title: Effects of Serial Skin Testing with Purified Protein Derivative on the Level and Quality of Antibodies to Complex and Defined Antigens in Mycobacterium bovis-Infected Cattle
Article Snippet: Paragraph title: Aerosol challenge procedures, mycobacterial culture, and assessment of lesions for experimental infection of cattle with Mycobacterium bovis . ... For the challenge inoculum, low-passage-number cultures (≤3 passages) of M. bovis were prepared, using standard techniques , in Middlebrook 7H9 liquid medium (Becton Dickinson, Franklin Lakes, NJ) supplemented with 10% oleic acid-albumin-dextrose complex (OADC) plus 0.05% Tween 80.

Derivative Assay:

Article Title: Two promoters in the esx-3 gene cluster of Mycobacterium smegmatis respond inversely to different iron concentrations in vitro
Article Snippet: Mycobacterium smegmatis mc2 155 (a gift from Rob Warren, South Africa) was used as the parent wildtype strain (WTms ) from which the MycP3 deletion mutant (ΔMycP3ms ) was derived, and as template to generate two complementation strains (ΔMycP3ms ::pr1MycP3ms and ΔMycP3ms ::pr2MycP3ms ). .. 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).

Cell Culture:

Article Title: Two promoters in the esx-3 gene cluster of Mycobacterium smegmatis respond inversely to different iron concentrations in vitro
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). .. Fe-free 7H9 (omitting ferric ammonium citrate) and Fe-free Sauton’s medium (3.5 mM KH2 PO4 (Sigma-Aldrich, USA, Catalogue No. NIST200B), 25 mM l -asparagine (Sigma-Aldrich, USA, Catalogue No. A0884), 10 mM citric acid (Sigma-Aldrich, USA, Catalogue No. 791725), 4 mM MgSO4 ·6H2 O (Sigma-Aldrich, USA, Catalogue No. 746452), 5% (v/v) glycerol, and 0.05% (v/v) Tween-80) were also used to monitor bacterial growth under iron-limiting condition.

Article Title: Region of Difference 2 Contributes to Virulence of Mycobacterium tuberculosis ▿
Article Snippet: .. Mycobacterium tuberculosis strains H37Rv, H37Rv:ΔRD2, H37Rv:ΔRD2::pMV306, H37Rv:ΔRD2::p7982, H37Rv:ΔRD2::p8586, H37Rv:ΔRD1, BCG Pasteur, and BCG Russia were cultured in Middlebrook 7H9 liquid medium (Becton Dickinson) containing 10% ADC (albumin dextrose complex; Becton Dickinson), 0.5% glycerol, and 0.08% Tween 80 (Sigma-Aldrich) or Middlebrook 7H10 agar containing 10% OADC (oleic acid plus ADC; Becton Dickinson), 0.5% glycerol, and PANTA (polymyxin B, amphotericin B, nalidixic acid, trimethoprim, and azlocillin; Becton Dickinson). .. Hygromycin or kanamycin (Sigma-Aldrich) was added to the medium when needed.

Digital PCR:

Article Title: The use of digital PCR to improve the application of quantitative molecular diagnostic methods for tuberculosis
Article Snippet: To assess reproducibility of dPCR measurements, when including the extraction step necessary for clinical analysis, two additional materials were used: A) 300 units of a whole mycobacterial panel in artificial sputum (BCG/ASM) were prepared. .. Mycobacterium bovis BCG strain Pasteur (BCG) was grown in 7H9 liquid medium (Becton-Dickinson (BD)) containing 10 % albumin-dextrose-catalase (ADC) (BD) enrichment medium and 0.2 % Tween 80.

other:

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 ▿ †
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.

Article Title: Antimicrobial Treatment Improves Mycobacterial Survival in Nonpermissive Growth Conditions
Article Snippet: Mycobacterium bovis BCG Glaxo strain and Mycobacterium tuberculosis H37Rv were grown in Sauton's or Middlebrook 7H9 liquid medium (Becton, Dickinson and Company) supplemented with albumin-dextrose complex.

Article Title: Mycobacterium bovis Induces Endoplasmic Reticulum Stress Mediated-Apoptosis by Activating IRF3 in a Murine Macrophage Cell Line
Article Snippet: M. bovis culture M. bovis Beijing strain was obtained from China Institute of Veterinary Drug Control (CVCC, China) and was grown in Middlebrook 7H9 liquid medium (BD Biosciences, NY, USA) with 10% ADC (albumin, dextrose, catalase), and 0.05% Tween-80 and grown to mid-log phase for 1 week at 37°C.

Polymerase Chain Reaction:

Article Title: Direct detection of Mycobacterium tuberculosis rifampin resistance in bio-safe stained sputum smears
Article Snippet: For culture, 45% of the suspension (135 μl) was inoculated into 7H9 liquid medium supplemented with Albumin Dextrose complex and PANTA (polymyxin B, amphotericin B, nalidixic acid, trimethoprim and azlocillin) supplement (Becton Dickinson, USA), and the tubes were incubated at 37°C for upto 8 weeks. .. The positivity in the 7H9 liquid culture was first identified on the basis of turbidity of the medium, which was then confirmed for presence of M . tuberculosis by ZN smear microscopy and IS6110 PCR (M . tuberculosis complex specific PCR) [ ].

Sonication:

Article Title: The use of digital PCR to improve the application of quantitative molecular diagnostic methods for tuberculosis
Article Snippet: The gDNA was initially quantified by fluorimetry using a dsDNA BR assay (Qubit, Thermo Fisher Scientific ), and was subsequently gravimetrically diluted to approximately 105 copies/μL in 1× TE pH 8.0 (Ambion) containing ~25 ng/μL sonicated human genomic DNA. .. Mycobacterium bovis BCG strain Pasteur (BCG) was grown in 7H9 liquid medium (Becton-Dickinson (BD)) containing 10 % albumin-dextrose-catalase (ADC) (BD) enrichment medium and 0.2 % Tween 80.

Binding Assay:

Article Title: The use of digital PCR to improve the application of quantitative molecular diagnostic methods for tuberculosis
Article Snippet: Two hundred units consisting of ~50 μL diluted gDNA were produced in low DNA binding tubes and stored at −20 °C. .. Mycobacterium bovis BCG strain Pasteur (BCG) was grown in 7H9 liquid medium (Becton-Dickinson (BD)) containing 10 % albumin-dextrose-catalase (ADC) (BD) enrichment medium and 0.2 % Tween 80.

Ziehl-Neelsen Stain:

Article Title: Increasing Prevalence of Nontuberculous Mycobacteria in Respiratory Specimens from US-Affiliated Pacific Island Jurisdictions 1
Article Snippet: A portion of the sediment was then used to prepare an AFB smear. .. The remaining sediment was resuspended and used to inoculate 7H9 liquid medium (BACTEC MGIT 960; Becton Dickinson, Franklin Lakes, NJ, USA), broth medium, and solid medium (Middlebrook 7H10 agar slant; Becton Dickinson).

DNA Extraction:

Article Title: Direct detection of Mycobacterium tuberculosis rifampin resistance in bio-safe stained sputum smears
Article Snippet: Paragraph title: Smear microscopy, M . tuberculosis culture and DNA isolation ... For culture, 45% of the suspension (135 μl) was inoculated into 7H9 liquid medium supplemented with Albumin Dextrose complex and PANTA (polymyxin B, amphotericin B, nalidixic acid, trimethoprim and azlocillin) supplement (Becton Dickinson, USA), and the tubes were incubated at 37°C for upto 8 weeks.

Mutagenesis:

Article Title: Two promoters in the esx-3 gene cluster of Mycobacterium smegmatis respond inversely to different iron concentrations in vitro
Article Snippet: Mycobacterium smegmatis mc2 155 (a gift from Rob Warren, South Africa) was used as the parent wildtype strain (WTms ) from which the MycP3 deletion mutant (ΔMycP3ms ) was derived, and as template to generate two complementation strains (ΔMycP3ms ::pr1MycP3ms and ΔMycP3ms ::pr2MycP3ms ). .. 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).

Microscopy:

Article Title: Direct detection of Mycobacterium tuberculosis rifampin resistance in bio-safe stained sputum smears
Article Snippet: Paragraph title: Smear microscopy, M . tuberculosis culture and DNA isolation ... For culture, 45% of the suspension (135 μl) was inoculated into 7H9 liquid medium supplemented with Albumin Dextrose complex and PANTA (polymyxin B, amphotericin B, nalidixic acid, trimethoprim and azlocillin) supplement (Becton Dickinson, USA), and the tubes were incubated at 37°C for upto 8 weeks.

Article Title: Increasing Prevalence of Nontuberculous Mycobacteria in Respiratory Specimens from US-Affiliated Pacific Island Jurisdictions 1
Article Snippet: The remaining sediment was resuspended and used to inoculate 7H9 liquid medium (BACTEC MGIT 960; Becton Dickinson, Franklin Lakes, NJ, USA), broth medium, and solid medium (Middlebrook 7H10 agar slant; Becton Dickinson). .. The prepared smear was heat-fixed at 70°C–80°C for 2 h. The smear was then stained by using the fluorochrome stain method, which used auramine O, a decolorizing solution, and a potassium permanganate counterstain and read by using a fluorescent microscope.

Mouse Assay:

Article Title: Mycobacterium tuberculosis from chronic murine infections that grows in liquid but not on solid medium
Article Snippet: Culture media and bacteria The media used were 7H9 liquid medium with 10% albumin, dextrose, catalase supplement and 0.05% Tween 80, and 7H11 agar medium with 10% oleic acid, albumin, dextrose, catalase supplement (Becton Dickinson, Oxford, UK). .. The two experiments both used 6-week, female Balbc mice, which were infected, with a mouse passaged H37Rv strain of M. tuberculosis suspended in 0.1% gelatin.

Lysis:

Article Title: Direct detection of Mycobacterium tuberculosis rifampin resistance in bio-safe stained sputum smears
Article Snippet: For culture, 45% of the suspension (135 μl) was inoculated into 7H9 liquid medium supplemented with Albumin Dextrose complex and PANTA (polymyxin B, amphotericin B, nalidixic acid, trimethoprim and azlocillin) supplement (Becton Dickinson, USA), and the tubes were incubated at 37°C for upto 8 weeks. .. For DNA isolation, the remaining 45% of the suspension (135 μl) was pelleted and re-suspended in 250 μl lysis solution (10% Chelex-100, 0.03% Triton X-100 and 0.3% Tween-20), heated at 90°C for 40 min., and centrifuged at ~8800×g for 10 min.

IA:

Article Title: Effects of Serial Skin Testing with Purified Protein Derivative on the Level and Quality of Antibodies to Complex and Defined Antigens in Mycobacterium bovis-Infected Cattle
Article Snippet: For the challenge inoculum, low-passage-number cultures (≤3 passages) of M. bovis were prepared, using standard techniques , in Middlebrook 7H9 liquid medium (Becton Dickinson, Franklin Lakes, NJ) supplemented with 10% oleic acid-albumin-dextrose complex (OADC) plus 0.05% Tween 80. .. Holstein steers were obtained from tuberculosis-free and paratuberculosis-monitored herds in Iowa and were housed in a biosafety level 3 (BSL-3) facility at the National Animal Disease Center (Ames, IA), according to institutional biosafety (permit IBC-0004RA) and animal care and use committee guidelines (with ethical approval via animal care and use protocol ACUP-3859).

Plasmid Preparation:

Article Title: The use of digital PCR to improve the application of quantitative molecular diagnostic methods for tuberculosis
Article Snippet: Two hundred × 50 μL units of the diluted plasmid were added into low DNA binding microcentrifuge tubes and stored at −20 °C. .. Mycobacterium bovis BCG strain Pasteur (BCG) was grown in 7H9 liquid medium (Becton-Dickinson (BD)) containing 10 % albumin-dextrose-catalase (ADC) (BD) enrichment medium and 0.2 % Tween 80.

Article Title: Two promoters in the esx-3 gene cluster of Mycobacterium smegmatis respond inversely to different iron concentrations in vitro
Article Snippet: Paragraph title: Bacterial strains, culture media and plasmid DNA ... 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).

Recombinant:

Article Title: Two promoters in the esx-3 gene cluster of Mycobacterium smegmatis respond inversely to different iron concentrations in vitro
Article Snippet: Bacterial strains, culture media and plasmid DNA Escherichia coli XL-1 blue (Stratagene, USA, Catalogue No. 200249) was used for manipulating and propagating recombinant plasmid DNA. .. 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).

Produced:

Article Title: The use of digital PCR to improve the application of quantitative molecular diagnostic methods for tuberculosis
Article Snippet: Two hundred units consisting of ~50 μL diluted gDNA were produced in low DNA binding tubes and stored at −20 °C. .. Mycobacterium bovis BCG strain Pasteur (BCG) was grown in 7H9 liquid medium (Becton-Dickinson (BD)) containing 10 % albumin-dextrose-catalase (ADC) (BD) enrichment medium and 0.2 % Tween 80.

Concentration Assay:

Article Title: Two promoters in the esx-3 gene cluster of Mycobacterium smegmatis respond inversely to different iron concentrations in vitro
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). .. Additionally, the iron deprivation rescuing of the M. smegmatis cultured in iron-deprived 7H9 or Sauton’s media was achieved by supplementation of ferric ammonium citrate in the same concentration of normal 7H9 or Sauton’s media.

Staining:

Article Title: Direct detection of Mycobacterium tuberculosis rifampin resistance in bio-safe stained sputum smears
Article Snippet: All smear slides were stained with ZN stain, observed and graded as per the Revised National Tuberculosis Control Programme (RNTCP) guidelines [ ]. .. For culture, 45% of the suspension (135 μl) was inoculated into 7H9 liquid medium supplemented with Albumin Dextrose complex and PANTA (polymyxin B, amphotericin B, nalidixic acid, trimethoprim and azlocillin) supplement (Becton Dickinson, USA), and the tubes were incubated at 37°C for upto 8 weeks.

Article Title: Increasing Prevalence of Nontuberculous Mycobacteria in Respiratory Specimens from US-Affiliated Pacific Island Jurisdictions 1
Article Snippet: The remaining sediment was resuspended and used to inoculate 7H9 liquid medium (BACTEC MGIT 960; Becton Dickinson, Franklin Lakes, NJ, USA), broth medium, and solid medium (Middlebrook 7H10 agar slant; Becton Dickinson). .. The prepared smear was heat-fixed at 70°C–80°C for 2 h. The smear was then stained by using the fluorochrome stain method, which used auramine O, a decolorizing solution, and a potassium permanganate counterstain and read by using a fluorescent microscope.

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    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
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    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

    RaaS is important for persistence of M. tuberculosis in prolonged stationary phase and in human macrophages. (A) Wild-type, Δ raaS Mtb , and Δ raaS Mtb com strains were incubated in 7H9 medium without shaking at 37°C for 7 months. **, Δ raaS Mtb had a significant survival defect ( P

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Antimicrobial Treatment Improves Mycobacterial Survival in Nonpermissive Growth Conditions

    doi: 10.1128/AAC.02774-13

    Figure Lengend Snippet: RaaS is important for persistence of M. tuberculosis in prolonged stationary phase and in human macrophages. (A) Wild-type, Δ raaS Mtb , and Δ raaS Mtb com strains were incubated in 7H9 medium without shaking at 37°C for 7 months. **, Δ raaS Mtb had a significant survival defect ( P

    Article Snippet: Mycobacterium bovis BCG Glaxo strain and Mycobacterium tuberculosis H37Rv were grown in Sauton's or Middlebrook 7H9 liquid medium (Becton, Dickinson and Company) supplemented with albumin-dextrose complex.

    Techniques: Incubation

    Volatiles released by M. tuberculosis grown in a 7H9 broth liquid medium.

    Journal: Beilstein Journal of Organic Chemistry

    Article Title: The volatiles of pathogenic and nonpathogenic mycobacteria and related bacteria

    doi: 10.3762/bjoc.8.31

    Figure Lengend Snippet: Volatiles released by M. tuberculosis grown in a 7H9 broth liquid medium.

    Article Snippet: Bacterial colonies from a 7H11 solid medium (Becton, Dickinson & Co., Sparks, USA, see ) or bacteria culture (100 µL) from a 7H9 liquid medium (Becton, Dickinson & Co., Sparks, USA, see ) were inoculated on a 7H11 solid medium and were spread out with sterile disposable loops to cover the entire plate/medium surface.

    Techniques: