Moxifloxacin, isoniazid, ethambutol, linezolid, clarithromycin, rifampicin, tyloxapol, and Tween80 were purchased from Sigma–Aldrich. Middlebrook 7H9 and Middlebrook 7H10 were purchased from Becton Dickinson. PBS was purchased from Invitrogen, Life Technologies (14080055).
Middlebrook 7h10
Manufactured by BD
Sourced in United States, France
Middlebrook 7H10 is a selective and enriched solid culture medium used for the isolation and cultivation of Mycobacterium species, particularly Mycobacterium tuberculosis. It provides essential nutrients and growth factors required for the optimal growth of these microorganisms.
Automatically generated - may contain errors
Lab products found in correlation
Middlebrook 7h9, by BD (10 mentions)
Tween 80, by Merck Group (4 mentions)
Middlebrook 7h9 broth, by BD (3 mentions)
Middlebrook 7h9 medium, by BD (3 mentions)
Rifampicin, by Merck Group (2 mentions)
Isoniazid, by Merck Group (2 mentions)
Ethambutol, by Merck Group (2 mentions)
Kanamycin, by Merck Group (2 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
Tyloxapol, by Merck Group (1 mentions)
Moxifloxacin, by Merck Group (1 mentions)
Linezolid, by Merck Group (1 mentions)
Clarithromycin, by Merck Group (1 mentions)
Inhalation exposure system model a4224, by Glas-Col (1 mentions)
Middlebrook oadc enrichment medium, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Merck Group (1 mentions)
Cell density meter, by Thermo Fisher Scientific (1 mentions)
3 mm sterile glass beads, by Merck Group (1 mentions)
Hygromycin, by Merck Group (1 mentions)
Oleic acid albumin dextrose catalase enrichment, by BD (1 mentions)
33 protocols using middlebrook 7h10
1
Mycobacterium Growth Inhibition Assay
2
Murine Model of Latent TB Infection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
M. tuberculosis H37Rv was grown in Middlebrook 7H9 broth (Becton, Dickinson and Company, Le Pont de Claix, France) supplemented with 10% Middlebrook OADC enrichment medium (Life Technologies, Gaitherburg, MD), 0.5% glycerol and 0.05% Tween 80 at 37 °C until log phase. H37Rv-GFP (kindly provided by Joel Ernst, University of San Fransciso, California) was grown similarly with 25 mg/L kanamycin in the broth. Prior to use, mycobacterial aliquots were passed 30× through a 29.5G needle to minimize bacterial clumping. Pulmonary infection at a dose of 50–100 live M. tuberculosis H37Rv bacilli was performed using a Glas-Col Inhalation Exposure System, Model A4224. Inoculum dosage was confirmed 24 h post-infection by determining the bacilli burden in the lungs of infected mice. To establish a latent infection, mice were treated with 25 mg/Kg INH-RIF for 6 weeks commencing at day 28 post- infection, which resulted in undetectable levels of bacilli. For disease reactivation, chemotherapy was withdrawn and bacilli burdens determined at day 90, 180 and 300 days post infection. For colony enumeration organs were weighed and homogenized in 0.04% Tween 80/PBS. Tenfold serial dilutions of organ homogenates were plated in duplicates on Middlebrook 7H10 (Becton, Dickinson and Company) agar plates containing 10% OADC (Life Technologies, Gaitherburg, MD) and incubated at 37 °C for 19–21 days.
3
Mycobacterial Susceptibility Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mycobacterial strains
were cultured in standard 7H9 media (Middlebrook 7H9 containing 0.2%
glycerol, 10% OADC, and 0.05% Tween-80) and diluted to a final CFU/mL
of ∼5 × 105 cells/mL. CFU totals were confirmed
to be within the range of 3–8 × 105 cells/mL
by plating dilutions of the inoculum on Middlebrook 7H10 (Becton Dickenson)
agar containing 0.5% glycerol and 10% OADC. Susceptibility was assessed
at 2 days (M. smegmatis, M. fortuitum, M. abscessus) or 7 days (M. tuberculosis H37Ra, M. bovis, M. avium). Cultures were incubated with 30 μg/mL resazurin for one
additional day to identify any effect on growth where compound solubility
was limiting. Reported data are the averages from at least two experiments.
were cultured in standard 7H9 media (Middlebrook 7H9 containing 0.2%
glycerol, 10% OADC, and 0.05% Tween-80) and diluted to a final CFU/mL
of ∼5 × 105 cells/mL. CFU totals were confirmed
to be within the range of 3–8 × 105 cells/mL
by plating dilutions of the inoculum on Middlebrook 7H10 (Becton Dickenson)
agar containing 0.5% glycerol and 10% OADC. Susceptibility was assessed
at 2 days (M. smegmatis, M. fortuitum, M. abscessus) or 7 days (M. tuberculosis H37Ra, M. bovis, M. avium). Cultures were incubated with 30 μg/mL resazurin for one
additional day to identify any effect on growth where compound solubility
was limiting. Reported data are the averages from at least two experiments.
4
Mycobacterial DNA Extraction Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Samples from Swiss TPH were incubated in liquid media Middlebrook 7H9 (Becton Dickinson) at 37 °C during two weeks, while as those from our laboratory were grown in commercial Middlebrook 7H10 agar (Becton Dickinson) with OADC supplement. In all cases, DNA extraction method was performed following the CTAB method31 . Purified DNA was used to perform our molecular approaches.
5
Antibiotic-Infused Middlebrook 7H10 Media
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Middlebrook 7H10 (Becton Dickinson, France) media containing the four first line anti-TB drugs, namely, rifampicin (R), isoniazid (H), streptomycin (S), and ethambutol (E) with the final concentration of 1 μg/mL, 0.2 μg/mL, 2 μg/mL, and 5 μg/mL (Sigma, St. Louis, USA) were prepared as recommended by the manufacturer, respectively.
6
Comparative Analysis of Mycobacterium Isolates
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
This study was carried out with 26 isolates belonging to INNO-LiPa cluster CHII recovered from clinical and environmental specimens by Prof. Françoise Portaels (Institute of Tropical Medicine, Antwerp, Belgium) and Prof. Roland Schulze-Röbbecke (University of Dusseldorf, Dusseldorf, Germany). M. smegmatis mc2155, M. tuberculosis H37Rv and the type strains of M. chelonae-M. abscessus complex (M. abscessus subsp. abscessus ATCC 19977T, M. abscessus subsp. bolletii CCUG 50184T, M. abscessus subsp. massiliense CCUG 48898T, M. chelonae ATCC 35752T, M. immunogenum ATCC 700505T, M. salmoniphilum ATCC 13758T, M. franklinii DSM 45524T, and M. saopaulense CCUG 66554T) were included for comparison (Table 1 ).
Cultures were grown aerobically at 28–30°C on solid media including Löwenstein-Jensen (LJ) and Middlebrook 7H10 [Becton-Dickinson (BD), USA] supplemented with oleic acid, albumin, dextrose and catalase (OADC–BD) and in liquid media including Middlebrook 7H9 (BD), Mueller-Hinton and Lisogeny Broth with 1% Tween 80.
Cultures were grown aerobically at 28–30°C on solid media including Löwenstein-Jensen (LJ) and Middlebrook 7H10 [Becton-Dickinson (BD), USA] supplemented with oleic acid, albumin, dextrose and catalase (OADC–BD) and in liquid media including Middlebrook 7H9 (BD), Mueller-Hinton and Lisogeny Broth with 1% Tween 80.
7
Quantifying Mycobacterial Burdens in Infected Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bacterial burdens in the brains, lungs, and spleens of infected mice were determined at specific time points after infection with M. tuberculosis. Organs were weighed and homogenized in 0.04% Tween 80 saline. Also, 10-fold serial dilutions of organ homogenates were plated in duplicates on Middlebrook 7H10 (Becton, Dickinson and Company) agar containing 10% OADC (Life Technologies, Gaitherburg, MD, USA), and incubated at 37°C for 19–21 days. The concentration of M. tuberculosis was then determined by counting the CFUs.
8
Standardized M. tuberculosis Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
M. tuberculosis Beijing strain was cultured in Middlebrook 7H10 (Becton Dickinson, Le Pont de Claix, France) supplemented with 10% oleic acid-albumin-dextrose catalase (OADC) (Becton Dickinson) for 30 days. The colonies were suspended in PBS, vigorously vortexed for 10 min using 3 mm sterile glass beads (Sigma-Aldrich, Saint-Quentin- Fallavier, France) and passed 10 times through a 25 G needle to disperse the clustered cells. The mycobacterial suspensions were then calibrated at 107 CFU/mL using an optical density of 580 nm (Cell Density Meter; Fisher Scientific, Illkirch, France) and were confirmed by counting the mycobacteria after Ziehl-Neelsen staining.
9
Quantifying Bacterial Burdens in Infected Organs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bacterial burdens in the brains, lungs and spleens of infected mice were determined at specific time points after infection with M. tuberculosis. Organs were weighed and homogenised in 0.04 % Tween 80 saline. Tenfold serial dilutions of organ homogenates were plated in duplicates on Middlebrook 7H10 (Becton, Dickinson and Company) agar plates containing 10 % OADC (Life Technologies, Gaithersburg, MD) and were incubated at 37 °C for 19–21 days. The concentration of M. tuberculosis was then determined by counting the CFUs.
10
Growth Conditions for Common Lab Organisms
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
M. tuberculosis H37Rv was grown on Middlebrook 7H10 agar (Becton Dickinson) supplemented with 10% vol/vol oleic acid, albumin, dextrose, and catalase (OADC) (Becton Dickinson) or Middlebrook 7H9 with (Becton Dickinson) OADC and 0.05% wt/vol Tween 80 (7H9-OADC-Tw). M. tuberculosis H37Rv constitutively expressing a red fluorescent protein (DsRed) (Carroll et al., 2018 (link)) was grown with 50 μg/mL hygromycin B. Staphylococcus aureus RN4220 and Escherichia coli DH5α were grown on LB agar. Mycobacterium smegmatis mc2155 was grown on 7H10 with 10% OADC. Pseudomonas aeruginosa BAA 47 was grown on tryptic soy agar, Saccharomyces cerevisiae Y187 was grown on YPD agar, and Bacillus subtilis Marburg was grown on nutrient agar.
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!