Bacteroides fragilis strain ATCC25285 was purchased from American Type Culture Collection (ATCC; Manassas, VA, USA). The strain was cultured in Brain Heart Infusion (BHI) broth anaerobically at 37 °C in anaerobic environment containing 5% H2, 10% CO2 and 85% N2 for 48 h. BHI was supplemented with 0.0005% hemin and 0.5 mg/mL vitamin K1 for optimal growth (BHIS)53 B. fragilis was centrifuged, washed three times with sterile PBS solution, and then was adjusted to 109 CFU/ml in sterile PBS through measuring the optical density at 600 nm using spectrophotometer (Thermo, USA). The culture supernatant of B. fragilis was filtered through a 0.25 μm filter and prepared for usage.
Atcc25285
Manufactured by American Type Culture Collection
Sourced in United States
ATCC25285 is a laboratory equipment item from the American Type Culture Collection. It is a sterile, single-use cell culture flask designed for the cultivation of various cell lines. The product provides a controlled environment for cell growth and experimentation.
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Lab products found in correlation
5 protocols using atcc25285
1
Culturing and Standardizing Bacteroides fragilis
2
Culturing and Standardizing Bacteroides fragilis
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Bacteroides fragilis strain ATCC25285 was purchased from American Type Culture Collection (ATCC; Manassas, VA, USA). The strain was cultured in Brain Heart Infusion (BHI) broth anaerobically at 37 °C in anaerobic environment containing 5% H2, 10% CO2 and 85% N2 for 48 h. BHI was supplemented with 0.0005% hemin and 0.5 mg/mL vitamin K1 for optimal growth (BHIS)53 B. fragilis was centrifuged, washed three times with sterile PBS solution, and then was adjusted to 109 CFU/ml in sterile PBS through measuring the optical density at 600 nm using spectrophotometer (Thermo, USA). The culture supernatant of B. fragilis was filtered through a 0.25 μm filter and prepared for usage.
3
Culturing and Standardizing Bacteroides fragilis
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Bacteroides fragilis strain ATCC25285 was purchased from American Type Culture Collection (ATCC; Manassas, VA, USA). The strain was cultured in Brain Heart Infusion (BHI) broth anaerobically at 37 °C in anaerobic environment containing 5% H2, 10% CO2 and 85% N2 for 48 h. BHI was supplemented with 0.0005% hemin and 0.5 mg/mL vitamin K1 for optimal growth (BHIS)53 B. fragilis was centrifuged, washed three times with sterile PBS solution, and then was adjusted to 109 CFU/ml in sterile PBS through measuring the optical density at 600 nm using spectrophotometer (Thermo, USA). The culture supernatant of B. fragilis was filtered through a 0.25 μm filter and prepared for usage.
4
Bacterial Growth and Filtration Protocol
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Bacteroides fragilis (B. fragilis) NTBF (Non Toxigenic Bacteroides fragilis), ATCC 25285 (Sears et al., 2014 (link); Choi et al., 2016 (link)), was purchased from the American Type Culture Collection. Bacteroides fragilis growth conditions were described in a previous study (Vernay et al., 2020 (link)). The supernatant was filtered through a 0.22 μm-pore-size syringe filter as we described in Vernay et al., 2020 (link).
Salmonella Heidelberg B182 strain (S. Heidelberg) was isolated on LB (Luria-Bertani) agar and grown in LB medium overnight at 37°C. S. Heidelberg was then subcultured by dilution in LB medium followed by incubation for 90 min at 37°C (Le Bars et al., 2012 (link)).
Salmonella Heidelberg B182 strain (S. Heidelberg) was isolated on LB (Luria-Bertani) agar and grown in LB medium overnight at 37°C. S. Heidelberg was then subcultured by dilution in LB medium followed by incubation for 90 min at 37°C (Le Bars et al., 2012 (link)).
5
Strain Identification and Microbial Contamination Assessment
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Strain ZY-312 was isolated from the feces of a healthy infant, and was previously identified using 16S rRNA gene sequence analysis (Deng et al., 2016 (link)). B. fragilis strain ATCC 25285 (also known as NCTC 9343) was purchased from the American Type Culture Collection (ATCC). The culture conditions and materials were as described previously for strains ZY-312 and ATCC 25285 (Deng et al., 2016 (link)). For the microbial contamination experiment, E. coli CBSLAM00087, Staphylococcus aureus (S. aureus) CMCC(B) 26058, Salmonella enterica (S. enterica) serotype Paratyphi B CBSLAM00994, Pseudomonas aeruginosa (P. aeruginosa) CBSLAM00818, Candida albicans (C. albicans) CMCC(F)98001, and Clostridium sporogenes (C. sporogenes) CMCC(B) 64941 were obtained from the Academy of Military Medical Science, Beijing, China. All strains were verified by PCR amplification of 16S rRNA gene using the universal primers 27 F (5′-AGAGTTTGATCCTGGCTCAG-3′) and 1492 R (5′-GGTTACCTTGTTACGACTT-3′) (Lanes D–J). Amplified products were sequenced (Biomed).
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