The five standard strains (A. baumannii ATCC 19606, S. aureus ATCC 6538P, P. aeruginosa ATCC 27853, P. aeruginosa ATCC 29260, E. coli ATCC 25257) were purchased from the Bioresource Collection and Research Center (Hsinchu, Taiwan) and inoculated into 5.0 mL LB broth and cultivated in a 200 rpm 37 °C incubator for 12 to 16 h. A total of 50.0 μL strains solution (3 × 108 CFU/mL) was added to 5.0 mL LA soft agar, vortexed, and mixed evenly, then poured on the LA medium to form a double layer. We took 30.0 μL of crude and various extracts solution (0.1 g/mL) into a sterile 6 mm filter paper disk, with DMSO (dimethyl sulfoxide) as the negative control group and tetracycline (7.50 mg/mL) as the positive control group. All plates were incubated at 37 °C for 12–16 h, observed, and measured for the size of the disk inhibition zone (DIZ).
E coli
Manufactured by RIKEN BioResource Center
E. coli is a common laboratory strain of bacteria used in research. It is a Gram-negative, rod-shaped bacterium that is widely used as a model organism in microbiology and molecular biology. E. coli is a versatile and well-understood microorganism that can be easily cultivated and genetically modified, making it a valuable tool for scientific investigations.
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3 protocols using e coli
1
Antimicrobial Susceptibility Testing Protocol
2
Microbial Growth and Strain Culturing
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Microbial media and growth plates were prepared prior to growing
various microbe strains. Sterilized Trypticase Soy Broth (TSB) (Merck
Co., Ltd.) and de Man, Rogosa, Sharpe (MRS) (Merck Co., Ltd.) broth
were used for all microbe cultures used in this study. Similarly,
sterilized Trypticase Soy Agar (TSA) (Merck Co., Ltd.) and MRS agar
(Merck Co., Ltd.) were poured and solidified in 100 × 15 mm petri
dishes.
Cultures of L. crispatus (Bioresource Collection and Research Center, Hsinchu, Taiwan) were
grown in MRS broth; on the other hand, MSSA, S. aureus, S. epidermidis, E.
faecalis, S. agalactiae, S. pneumoniae, E.
coli, K. pneumoniae, and C. albicans (Bioresource Collection
and Research Center) were grown in TSB. All cultures were prepared
in a shaking incubator (Thermo Fisher Scientific) set at 37 °C
and 200 rpm for 24 h. The cultures were diluted to a 0.5 MacFarland
bacterial turbidity standard using a UV–VIS optical density
spectrophotometer (Vernier Software & Technology, Beaverton, OR,
USA). 50 μL of different microbes was added to their corresponding
nutrient plate, and sterilized cell spreaders were used to equally
distribute the microbes on the plate.
various microbe strains. Sterilized Trypticase Soy Broth (TSB) (Merck
Co., Ltd.) and de Man, Rogosa, Sharpe (MRS) (Merck Co., Ltd.) broth
were used for all microbe cultures used in this study. Similarly,
sterilized Trypticase Soy Agar (TSA) (Merck Co., Ltd.) and MRS agar
(Merck Co., Ltd.) were poured and solidified in 100 × 15 mm petri
dishes.
Cultures of L. crispatus (Bioresource Collection and Research Center, Hsinchu, Taiwan) were
grown in MRS broth; on the other hand, MSSA, S. aureus, S. epidermidis, E.
faecalis, S. agalactiae, S. pneumoniae, E.
coli, K. pneumoniae, and C. albicans (Bioresource Collection
and Research Center) were grown in TSB. All cultures were prepared
in a shaking incubator (Thermo Fisher Scientific) set at 37 °C
and 200 rpm for 24 h. The cultures were diluted to a 0.5 MacFarland
bacterial turbidity standard using a UV–VIS optical density
spectrophotometer (Vernier Software & Technology, Beaverton, OR,
USA). 50 μL of different microbes was added to their corresponding
nutrient plate, and sterilized cell spreaders were used to equally
distribute the microbes on the plate.
3
Cultivation and Maintenance of Pathogenic Bacteria
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P. aeruginosa (ATCC 27853), E. coli (ATCC 25922) and S. aureus (ATCC 6538) were obtained from Bioresource Collection and Research Center, Taiwan. Some experiments used biofilm-producing S. aureus SA 113 (ATCC 35556). Colonies were maintained on agar supplemented with the necessary medium, and transferred to fresh agar plates every two weeks. Cultures were grown by taking 2–3 colonies from agar plate to 25 mL of medium in a flask (3 mL of in tubes for some experiments) and maintaining at 37 °C at 180 rpm for 12–16 hours. Medium used for P. aeruginosa and S. aureus was DifcoTM nutrient broth. DifcoTM tryptic soy broth was for E. coli. On the other hand, S. aureus SA 113 required tryptic soy broth supplemented with 0.5% glucose.
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P. aeruginosa (ATCC 27853), E. coli (ATCC 25922) and S. aureus (ATCC 6538) were obtained from Bioresource Collection and Research Center, Taiwan. Some experiments used biofilm-producing S. aureus SA 113 (ATCC 35556). Colonies were maintained on agar supplemented with the necessary medium, and transferred to fresh agar plates every two weeks. Cultures were grown by taking 2–3 colonies from agar plate to 25 mL of medium in a flask (3 mL of in tubes for some experiments) and maintaining at 37 °C at 180 rpm for 12–16 hours. Medium used for P. aeruginosa and S. aureus was DifcoTM nutrient broth. DifcoTM tryptic soy broth was for E. coli. On the other hand, S. aureus SA 113 required tryptic soy broth supplemented with 0.5% glucose.
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