Anaerogen bags

Manufactured by Thermo Fisher Scientific

Sourced in France

AnaeroGen bags are lab equipment designed to create an anaerobic environment. They facilitate the generation of an oxygen-free atmosphere within a confined space for various applications that require an anaerobic environment.

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Lab products found in correlation

Generbox jars, by bioMérieux (2 mentions) Unimax 2010, by Heidolph (1 mentions) Double pore slim electrode, by Hamilton Company (1 mentions) Violet red bile agar, by Thermo Fisher Scientific (1 mentions) M17 broth, by Thermo Fisher Scientific (1 mentions)

4 protocols using anaerogen bags

Anaerobic and Aerobic Growth Assay

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All strains were screened in microplate (96-well) experiments for their ability to grow (16 and 42 h at 37°C) in a. anaerobiosis (AN; static cultivation in MRS broth in Generbox jars, bioMérieux SA, Marcy-l’Etoile, France, with AnaeroGen bags, Oxoid), b. aerobiosis (AE; in MRS broth, agitation on a rotary shaker at 150 rpm; Unimax 2010, Heidolph Instruments GmbH & Co.KG, Germany) and c. heme-supplemented aerobiosis (AEH; in MRS broth with 2.5 µg/mL hemin, initial pH 6.8, agitation on a rotary shaker at 150 rpm). Microplates (180 µL substrate/well) were inoculated with 20 µL of standardized (OD₄₅₀ = 2.0) overnight anaerobic MRS-pre-cultures. Optical density at 450 nm (OD₄₅₀; Titertek Multiskan Plus 311 BO Microplate Reader) and pH values (Double Pore Slim electrode, Hamilton Company, Reno, Nevada, USA) were measured at 16 h and 42 h on two replicates.
Production of catalase was qualitatively assayed by re-suspending the washed biomass (final OD₆₅₀ = 1.0) derived from 1 mL of AN, AE and AEH cultures (16 h, 37°C) in 100 µL of 3% (v/v) H₂O₂. Bubble formation provided an indication of the presence of catalase activity in cell suspensions [23] .

Zotta T., Ricciardi A., Ianniello R.G., Parente E., Reale A., Rossi F., Iacumin L., Comi G, & Coppola R. (2014). Assessment of Aerobic and Respiratory Growth in the Lactobacillus casei Group. PLoS ONE, 9(6), e99189.

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Pneumococcal Transformation by Competence

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S. pneumoniae was grown in liquid C medium (67 (link)) or on Todd-Hewitt (BD Difco) agar plates at 37°C. Plates were incubated in an anaerobic chamber made by including AnaeroGen bags from Oxoid in a sealed container. For selection of transformants, kanamycin or streptomycin was added to the growth medium to a final concentration of 400 or 200 μg/ml, respectively. Genetic transformation of S. pneumoniae was performed by natural competence as previously described by Straume et al. (68 (link)).

Alcorlo M., Straume D., Lutkenhaus J., Håvarstein L.S, & Hermoso J.A. (2020). Structural Characterization of the Essential Cell Division Protein FtsE and Its Interaction with FtsX in Streptococcus pneumoniae. mBio, 11(5), e01488-20.

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Microbial Enumeration in Cheese Making

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Microbial counts were performed on milk, milk after the addition of starter, curd before stretching, and on cheeses at 7, 15, and 30 d. Cheese samples were homogenized in 2% (wt/vol) trisodium citrate solution and further decimal dilutions were prepared in sterile quarter-strength Ringer's solution; all dilutions were carried out in Ringer's for milk samples. Total mesophilic counts were carried out by pour plating in standard plate count agar (PCA, Oxoid) after 48 h at 30°C. Thermophilic streptococci were enumerated in LM17 agar (M17 broth, Oxoid, with 1% lactose and 1.2% agar bacteriological) after incubation for 2 d at 42°C. Nonstarter lactic acid bacteria were differentially enumerated in modified MRS with bromophenol blue (mMRS-BPB; Lee and Lee, 2008) after incubation under anaerobiosis (GenBox Jars, bioMérieux Italia, Firenze, with AnaeroGen bags, Oxoid) at 25 or 37°C for 48 h. Coliforms were enumerated in milk and cheese by pour-plating on violet red bile agar (Oxoid), after incubation for 24 h at 37°C.

Guidone A., Braghieri A., Cioffi S., Claps S., Genovese F., Morone G., Napolitano F, & Parente E. (2015). Effect of adjuncts on microbiological and chemical properties of Scamorza cheese. Journal of dairy science, 98(3).

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Anaerobic Bacterial Characterization Protocol

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Overnight broth cultures were streaked on MRS medium modified according to Lee and Lee (2008) by adding 0.05 % w/v cysteine and 0.002 % w/v bromophenol blue, pH 6.5 (mMRS-BPB). Incubation was carried out in anaerobiosis (Generbox jars, bioMérieux SA, Marcy-l'Etoile, France, with AnaeroGen bags, Oxoid) for 48 h at the temperatures listed in paragraph 2.1. The set of binary characters used to score colony morphology is listed in Table 1.
Cell morphology was evaluated by phase contrast microscopy (1000×) on overnight cultures in modified APT broth (Sperber and Swan 1976) . Phase contrast microscopy was also used to evaluate CO 2 production via glucose using the hotloop technique (Sperber and Swan 1976) . Colony morphology was observed under white fluorescent light (220-240 V, 65 W, 60 lm/W, 3900 lm, Philips) under a laminar flow hood and the results were scored using a set of binary attributes (Table 1). Data were collected over several months by trained operators.
A combination of multivariate statistical methods (hierarchical cluster analysis, multidimensional scaling, classification trees) was used for the statistical treatment of the data using Systat 13 (Systat Inc., Chicago, IL, USA).

Ricciardi A., Parente E., Tramutola A., Guidone A., Ianniello R.G., Pavlidis D., Tsakalidou E, & Zotta T. (2015). Evaluation of a differential medium for the preliminary identification of members of the Lactobacillus plantarum and Lactobacillus casei groups. Annals of microbiology, 65(3).

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