Tetrathionate-based memory was induced under in vitro conditions in either liquid culture or during growth on plates. For liquid culture induction, strains were backdiluted 1:1000 from overnight culture in SOC broth into pre-reduced anaerobic SOC broth and grown at 37°C in an anaerobic chamber (Coy Lab Products) using 7%H2/ 20% CO2/63% N2 culture gas. After 1h, potassium tetrathionate (Sigma) dissolved in pre-reduced SOC media was added to cultures and induction was undertaken at 37°C in the anaerobic chamber for 4h. Memory was assayed by plating and growth in aerobic conditions of serial dilutions of the bacteria on luria-broth (LB) + 300μg/mL streptomycin sulfate (Sigma) + 60μg/mL 5-Bromo-4-chloro-3-indolyl b-D-galactopyranoside (x-gal) (Santa Cruz Biotechnology) agar plates. Switching levels were estimated through counting greater than 250 blue and white colonies unless otherwise noted. In vitro testing on solid plates involved plating on LB + 300μg/mL streptomycin sulfate (Sigma) + 60μg/mL x-gal (Santa Cruz Biosciences) + 10 mM sodium tetrathionate (Sigma) agar. Growth was maintained in anaerobic conditions for approximately 8–12 hours using the Gaspak system (BD Biosciences), followed by further growth in aerobic conditions to allow development of memory and the x-gal reporter.
Streptomycin sulfate
Manufactured by Merck Group
Sourced in United States, Germany, Italy, Spain, Macao, United Kingdom, Sao Tome and Principe, Canada, China, Israel
Streptomycin sulfate is a white to pale yellow, crystalline powder. It is a broad-spectrum antibiotic that is effective against various bacteria, including Gram-positive and Gram-negative species.
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Lab products found in correlation
Penicillin, by Merck Group (68 mentions)
Penicillin g, by Merck Group (59 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (58 mentions)
L glutamine, by Merck Group (20 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (17 mentions)
Gentamicin sulfate, by Merck Group (11 mentions)
Amphotericin b, by Merck Group (10 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (10 mentions)
Fetal bovine serum (fbs), by Merck Group (10 mentions)
Penicillin g sodium, by Merck Group (9 mentions)
Trypsin, by Merck Group (9 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (9 mentions)
Bovine serum albumin (bsa), by Merck Group (8 mentions)
Penicillin g sodium salt, by Merck Group (7 mentions)
Potassium penicillin, by Merck Group (7 mentions)
Streptomycin sulfate, by Thermo Fisher Scientific (7 mentions)
Kanamycin sulfate, by Merck Group (6 mentions)
Glutamine, by Merck Group (6 mentions)
Glutamax, by Thermo Fisher Scientific (6 mentions)
Ampicillin, by Merck Group (5 mentions)
255 protocols using streptomycin sulfate
1
Tetrathionate-Induced Bacterial Memory
2
Enterococcus faecalis Growth in Streptomycin
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A growth medium for streptomycin-resistant T2-strain Enterococcus faecalis (E. faecalis) (ATCC® 29212™, ATCC, Manassas, VA, USA) was prepared and autoclaved. To prevent contamination by additional bacterial species, 0.5 mg/mL streptomycin sulfate (Sigma-Aldrich Co.) was added, as E. faecalis is resistant to this concentration of streptomycin sulfate. Each tooth specimen was filled from the coronal part of the root canal with the freshly prepared bacterial suspension and then incubated at 37 °C and 100% humidity. The bacterial suspension was replaced with a fresh preparation every 24 h for 21 days.
3
Antifungal Activity of Streptomyces Strains
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For preparing culture filtrates for each selected Streptomyces spp., each strain was grown on 2% potato dextrose broth (PDB; Difco, Detroit, MI) by incubating at 30 °C with constant shaking at 140 rpm for 7 days. To decant the supernatant, suspensions of each Streptomyces spp. were centrifuged for 15 min at 9000 rpm. The supernatant was filtered with 0.4 μm nucleopore membranes. The filtered suspension was added to the media. Two types of media, heated and unheated media, were used for the assay to evaluate the antifungal activity against WDF. For the heated media (PHM), filtered suspensions of each strain were added after autoclave sterilization of PDA supplemented with 100 mg/L streptomycin sulfate (Sigma-Aldrich, Steinheim, Germany), whereas the filtered suspensions of each strain were directly added to PDA supplemented with 100 mg/L streptomycin sulfate without autoclaving for the unheated media (PUHM).
Agar plugs were excised from the growing edge of WDF isolates grown on PDA at 25 °C for 10 days and transferred onto either PHM or PUHM and incubated at 25 °C for 7 days to measure the rate of suppression (S) of the fungal growth. Controls were prepared by adding DW to PDA. The rate of suppression was calculated as below:
where C is the mycelial growth for control (PDA + DW), T is the mycelial growth for treatment (either PHM or PUHM).
Agar plugs were excised from the growing edge of WDF isolates grown on PDA at 25 °C for 10 days and transferred onto either PHM or PUHM and incubated at 25 °C for 7 days to measure the rate of suppression (S) of the fungal growth. Controls were prepared by adding DW to PDA. The rate of suppression was calculated as below:
where C is the mycelial growth for control (PDA + DW), T is the mycelial growth for treatment (either PHM or PUHM).
4
Resuscitation and Culture of Cervical Cancer Cell Lines
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The human cervical cancer cell lines SiHa and CaSki were obtained from the American Type Culture Collection (Manassas, VA, USA) and resuscitated by the Cell Bank of the Chinese Academy of Science (Shanghai, China). SiHa and CaSki cells were routinely cultured in minimum essential medium and RPMI-1640 medium, respectively, with 10% fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific, Inc.), 100 IU/ml penicillin G, and 100 mg/ml streptomycin sulfate (Sigma-Aldrich; Merck Millipore). The incubation conditions were 37°C under 5% CO2 and 95% air atmosphere at constant humidity. The culture medium was changed every other day.
5
Cell culture of RCC and 293 cell lines
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The human RCC cell lines 786-O and A498, and 293 cells were obtained from the American Type Culture Collection (Manassas, VA, USA). All cells were cultured in Dulbecco's modified Eagle's medium (DMEM; Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA), supplemented with 10% (v/v) fetal bovine serum (Gibco; Thermo Fisher Scientific), 1% 100 U/ml penicillin and 1% 100 mg/ml streptomycin sulfate (Sigma-Aldrich: Merck KGaA, Darmstadt, Germany) at 37°C in a humidified atmosphere containing 5% CO2.
6
Isolation of Canine Cumulus-Oocyte Complexes
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Ovaries were kept in saline solution (0.9%, w/v NaCl) containing 100 IU/mL of penicillin G and 0.1 mg/mL of streptomycin sulfate (Merck KGaA, Darmstadt, Germany), then transported to the laboratory within 20 min. During transport, the temperature of the ovaries was maintained at 37 °C. After washing in phosphate-buffered saline (PBS) (137 mM NaCl, 2.7 mM KCL, 10 mM Na2HPO4, 2 mM KH2PO4) at 38 °C and pH 7.4, the ovaries were mechanically dissected.
Only antral follicles from 0.5 mm to 4.9 mm (anestrus, proestrus and diestrus) or 5 to 10 mm (estrus) were punctured with a narrow bore Pasteur pipette, and cumulus–oocyte complexes (COCs) with more than three tightly wrapped layers of CCs and uniformly distributed oocyte cytoplasm were selected under a dissecting microscope (Meiji Techno SKT, Tokyo, Japan). The selected COCs retrieved from pooled antral follicles of individual bitches according to each reproductive phase were collected separately in Petri dishes containing PBS.
Only antral follicles from 0.5 mm to 4.9 mm (anestrus, proestrus and diestrus) or 5 to 10 mm (estrus) were punctured with a narrow bore Pasteur pipette, and cumulus–oocyte complexes (COCs) with more than three tightly wrapped layers of CCs and uniformly distributed oocyte cytoplasm were selected under a dissecting microscope (Meiji Techno SKT, Tokyo, Japan). The selected COCs retrieved from pooled antral follicles of individual bitches according to each reproductive phase were collected separately in Petri dishes containing PBS.
7
Critical Concentration of H2O2 for Saccharomyces boulardii
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The modified protocol was based on the use of a strain of Saccharomyces boulardii from the collection of the Laboratory of Pharmaceutical Biotechnologies, Faculty of Biotechnologies, USAMV (University of Agronomic Science and Veterinary Medicine) Bucharest. The viability analysis required to calculate the critical point was performed using peptone yeast extract agar (g/L; yeast extract 5g, streptomycin sulfate, 0.03 g, soya peptone, 10 g, glucose, 40 g, chloramphenicol, 0.05 g, agar, 15 g; Merck KGaA, Darmstadt, Germany). The cross between the viability and mortality lines (different concentrations of H2O2 (%)—0.1, 0.3, 0.5, 1, 2) defined the critical concentration, and was expressed as a critical point (%). The untreated sample represented the control. The antioxidant potential in vivo was expressed as percentage after the bioactive compound assimilation [26 (link)].
8
Iron-Based Hydrogel Synthesis
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All chemicals used in this study were reagent grade and were used as such without purification. Ferrous chloride tetrahydrate (FeCl2 · 4H2O) (>99%), ferric chloride hexahydrate (FeCl3 · 6H2O) (99%), and streptomycin sulfate were obtained from Merck KGaA (Darmstadt, Germany). Low molecular weight CS (deacetylation 75%–85%), PEG, KA (>98% purity; molecular weight 142.11 g/mol), and ampicillin were purchased from Sigma-Aldrich (Saint Louis, MO, USA) and were used as such without any pretreatment. Acetic acid (99.8%) was purchased from Hamburg Industries, Inc (Hamburg, Germany). Deionized water was used for all the experiments.
9
Isolation and Quantification of Pithomyces chartarum in Grass Samples
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Samples consisting of approximately 500 to 700 g of fresh grass were collected from 4 different locations in the sheep grazing areas. Selected plant materials were investigated to detect P. chartarum in them. First, plant fragments were surface sterilized (ethanol 75% 0.5 min, sodium hypochlorite 3% 1 min, and 4 to 5 washes in autoclaved bi-distilled water), placed in Petri plates containing potato dextrose agar (Panreac, Spain), and medium supplemented with streptomycin sulfate (0.5 g/L; Merck, Germany) to avoid bacterial contaminations. They were incubated 3 to 5 days at 26°C in the dark to obtain endophytic isolates of P. chartarum. The second procedure was to determine the presence of the level of conidia in the grass. With this objective, sterile double-distilled water was passed through funnels containing approximately 50 gm of plant material selected from the prospected samples. The eluent was collected and subsequently analyzed, quantifying the presence of P. chartarum conidia under the optical microscope with the aid of a hemocytometer.
10
Isolation of T. cruzi Trypomastigotes from Vero Cells
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Y T. cruzi strain was obtained from Dr. David (Harvard Medical School, Boston, MA, USA) and VFRA strain from ChagasEpiNet VII Framework Consortium. Vero cells (African green monkey kidney, ATCC) were grown in RPMI medium supplemented with 5% fetal bovine serum (FBS), 100 UI/mL of antibiotics mixture, 10 μg/mL streptomycin (Streptomycin Sulfate BioChemica, PanReac AppliChem) and 2 mM L-glutamine (Merck) at 37 °C in an atmosphere of 5% CO 2 until the cells reached 80% confluence. The cell monolayer was subsequently infected with metacyclic trypomastigotes (VFRA and Y, 10 parasites per cell). After 4 days, the supernatant medium was collected. Trypomastigotes were obtained from these Vero cell co-cultures by differential centrifugation at 1000 g for 5 min to eliminate the amastigotes and dead cells and centrifugation at 1600 g for 10 min to isolate them.
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