Djulis (Chenopodium formosanum) was purchased from Taitung country farmers’ association (Taitung, Taiwan). Wheat flour was purchased from Yifeng food company (New Taipei City, Taiwan). MRS broth was purchased from Hardy Diagnostics (Santa Maria, CA, USA). Peptone and Bacto agar were purchased from Bioshop (Burlington, ON, Canada). Glucose, sucrose, lactose, magnesium sulfate, sodium phosphate dibasic, potassium phosphate dibasic 2-mercaptoethanol, sodium dodecyl sulfate, sodium tetraborate, monosodium dihydrogen orthophosphate, sodium phosphate dibasic, 2,2-diphenyl-1-picrylhydrazyl, sodium chloride, sodium carbonate, potassium persulfate, hydrochloric acid, 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid), trolox (6-hydroxy-2,5,7,8-tetramethylchromane-2-carboxylic acid) were purchased from Sigma-Aldrich (St. Louis, MO, USA). o-Phthaldialdehyde was purchased from Merck (Darmstadt, Germany).
Mrs broth
Manufactured by Hardy Diagnostics
Sourced in United States
MRS broth is a culture medium used for the growth and enumeration of lactic acid bacteria, particularly lactobacilli. It provides the essential nutrients and growth factors required by these microorganisms. The broth is formulated according to the original recipe published by de Man, Rogosa, and Sharpe.
Automatically generated - may contain errors
Lab products found in correlation
Monosodium dihydrogen orthophosphate, by Merck Group (1 mentions)
Potassium persulfate, by Merck Group (1 mentions)
Magnesium sulfate, by Merck Group (1 mentions)
2 2 azino bis 3 ethylbenzothiazoline 6 sulphonic acid, by Merck Group (1 mentions)
6 hydroxy 2 5 7 8 tetramethylchromane 2 carboxylic acid trolox, by Merck Group (1 mentions)
O phthaldialdehyde, by Merck Group (1 mentions)
Sodium carbonate, by Merck Group (1 mentions)
Sodium dodecyl sulfate (sds), by Merck Group (1 mentions)
Hydrochloric acid (hcl), by Merck Group (1 mentions)
Sucrose, by Merck Group (1 mentions)
Sodium phosphate dibasic, by Merck Group (1 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (1 mentions)
Lactose, by Merck Group (1 mentions)
Sodium tetraborate, by Merck Group (1 mentions)
Ribose, by Merck Group (1 mentions)
Ammonium citrate, by Merck Group (1 mentions)
Beef extract, by BD (1 mentions)
Sodium lactate, by Merck Group (1 mentions)
Xylose, by Merck Group (1 mentions)
Yeast extract, by BD (1 mentions)
5 protocols using mrs broth
1
Chenopodium formosanum Cultivation Protocol
2
Culturing and Quantifying L. reuteri
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L. reuteri bacteria were grown on deMan, Rogosa and Sharpe (MRS) agar (Hardy Diagnostics, Santa Maria, CA, USA) for 24 h at 37 °C. The bacteria were then transferred to an MRS broth (Hardy Diagnostics) and quantified by serial dilution plating and counting colonies. Bacteria were diluted to 1010 or 109 CFU/mL for storage, aliquoted and frozen at −80 °C. For all assays, bacterial aliquots were thawed at room temperature, centrifuged (6000× g, 2 min), rinsed with cold sterile Hank’s Balanced Salt Solution (HBSS; containing 1.0 g/L glucose, 0.011 g/L phenol red and 0.35 g/L sodium bicarbonate, pH 7.1–7.5; H9269; Sigma-Aldrich) and then resuspended in HBSS and diluted to the concentrations used in the respective assays.
The viability of the bacteria under standard experimental conditions (108 CFU/mL live bacteria in HBSS, 100 µM KYN, 3 h incubation) was evaluated by serial dilution plating and counting colonies.
The viability of the bacteria under standard experimental conditions (108 CFU/mL live bacteria in HBSS, 100 µM KYN, 3 h incubation) was evaluated by serial dilution plating and counting colonies.
3
Anaerobic Growth of Gut Microbes
4
Determining Paenibacillus wasatchensis Growth Substrates
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DeMan, Rogosa, and Sharpe (MRS) broth (Hardy Diagnostics) supplemented with 1% ribose (Sigma Aldrich Inc.) was used to prepare working cultures of Pa. wasatchensis by incubation at 30°C for 4 d. A carbohydrate-restricted MRS (CR-MRS) was also prepared as described previously (McMahon et al., 2020) (link) using 10 g of protease peptone No. 3 (EMD Chemicals Inc.), 10 g of beef extract (Becton Dickinson), 5 g of yeast extract (Becton Dickinson), 5.0 g of sodium acetate (Sigma Aldrich Inc.), 1.0 g of Tween-80, 2.0 g of ammonium citrate (Sigma Aldrich Inc.), 2.0 g of dipotassium phosphate (Fisher Scientific), 0.1 g of magnesium sulfate (Fisher Scientific), and 0.05 g of manganese sulfate (Avantor Performance Materials Inc.) made up to 1 L using distilled water. This CR-MRS plus 0.5% of either sodium gluconate (Tokyo Chemical Industry Ltd.), sodium lactate (Sigma Aldrich Inc.), xylose (Sigma Aldrich Inc.), or ribose was used for determining the growth of Pa. wasatchensis strains in different substrates. For gas production studies, CR-MRS plus 1% mixtures containing gluconate and ribose at 100:0, 70:30, 50:50, and 30:70 ratios were used. For studying gas production with gluconate as the only substrate, CR-MRS plus 0.4% to 1.0% gluconate was used.
5
Isolation and Cultivation of Lactobacillus Strains
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Various Lactobacillus species were isolated from commercial yogurts, including Suki, Oikos, and Yoplait yogurt purchased from the supermarket (Table 1). Lactobacillus rhamnosus was purchased from ATCC (LGG, ATCC# 53103, Manassas, VA). All Lactobacilli were grown in the De Man, Rogosa, and Sharpe (MRS) broth (Hardy Diagnostics, Santa Maria, CA) at 37°C and incubated in a BactronEZ SHEL LAB anaerobic chamber (Sheldon Manufacturing, Inc. Cornelius, OR). MRS agar plates were made with 55g Lactobacilli MRS broth of dehydrated culture media and 15 g agar (Fisher Scienti c) in one liter of water. The lysogeny broth (LB) medium and agar plates for Escherichia coli (E. coli) growth were purchased from Criterion and prepared according to the manufacturer's instructions. Mice BALB/c mice (6-8 weeks of age) were purchased from Jackson Laboratory. Mice were housed and fed in a speci c pathogen-free animal house. All animal experiments were approved by the Institutional Animal Care and Use Committee of Nanjing Medical University.
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