Pyrolyzing citric acid was used to synthesize GO. First, 1 gr of citric acid (Sigma-Aldrich, St. Louis, MO, USA) was put into a 5 mL beaker and heated to 200 °C. After 5 min, the citric acid was liquefied and the color changed from colorless to yellow. After 30 min, the color changed to orange. The heating was kept until it changed into black liquid in 100 min, suggesting the formation of GO. The black liquid was then added to KH2PO4 solution (Sigma-Aldrich, St. Louis, MO, USA) and by 80 mg/mL KOH solution to obtain 1.55 M KH2PO4 solution 0.8 wt% GO with pH 6.5. [29 (link),30 (link)]. The solution was stored at 4 °C before use.
Kh2po4 solution
Manufactured by Merck Group
Sourced in United States
KH2PO4 solution is a laboratory reagent used as a buffer solution to maintain a specific pH range in various applications. It is a clear, colorless liquid composed of potassium dihydrogen phosphate dissolved in water. The solution is commonly used in biochemical, analytical, and cell culture procedures to control the acidity or basicity of the environment.
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Lab products found in correlation
4 protocols using kh2po4 solution
1
Synthesis of Graphene Oxide from Citric Acid
2
Quantitative Analysis of Phosphate Solubilization
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A quantitative analysis of P solubilization activity was performed using the molybdate blue color method (59 (link)). Briefly, bacterial isolates were inoculated in 25 ml Pikovskaya broth medium (31 ) and incubated for 7 days at 28°C with shaking at 150 rpm. Bacterial cultures were centrifuged at 15,000 rpm for 30 min. The supernatant (1 ml) was mixed with 10 ml of chloromolybidic acid and the volume was made up to 45 ml with distilled water. Cholorostannous acid (0.25 ml) was added, and the volume was brought up to 50 ml with distilled water. The absorbance of the developing blue color was measured by spectrophotometry (Thermo, USA) at 600 nm. The amount of solubilized phosphate was detected using a standard curve produced with dilutions of a KH2PO4 solution (Sigma-Aldrich, USA).
3
Quantitative Analysis of Phosphate Solubilization
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A quantitative analysis of P solubilization activity was performed using the molybdate blue color method (59 (link)). Briefly, bacterial isolates were inoculated in 25 ml Pikovskaya broth medium (31 ) and incubated for 7 days at 28°C with shaking at 150 rpm. Bacterial cultures were centrifuged at 15,000 rpm for 30 min. The supernatant (1 ml) was mixed with 10 ml of chloromolybidic acid and the volume was made up to 45 ml with distilled water. Cholorostannous acid (0.25 ml) was added, and the volume was brought up to 50 ml with distilled water. The absorbance of the developing blue color was measured by spectrophotometry (Thermo, USA) at 600 nm. The amount of solubilized phosphate was detected using a standard curve produced with dilutions of a KH2PO4 solution (Sigma-Aldrich, USA).
4
Screening Phosphate Solubilization Capacity
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Phosphate solubilization by the isolated fluorescent Pseudomonas isolates was tested in solid media by the method described by Nautiyal70 (link). Ten microliters of the culture of each strain was spotted on the surface of the solid media in Petri dishes. The solubilization capacity was assessed by the transparent area formed around the colony. Ten days after incubation at 30 °C, the diameter of the solubilization halo (DSH) was determined by the following formula (Eq. 1 ): where THD is the total halo diameter, and CD is the colony diameter.
Quantitative estimation of phosphate solubilization in broth was carried out in Erlenmeyer flasks (250 ml) containing 100 ml of NBRIP medium70 (link). The flasks were incubated at 28 °C for 5 days at 120 rpm. Then, 5 ml of each isolate culture was centrifuged at 3000 rpm for 20 min. Phosphate concentration in culture supernatant was estimated as described by Olsen and Sommers71 . Three replicates were used, and a standard calibration curve was made with KH2PO4 solution (Sigma-Aldrich).
Quantitative estimation of phosphate solubilization in broth was carried out in Erlenmeyer flasks (250 ml) containing 100 ml of NBRIP medium70 (link). The flasks were incubated at 28 °C for 5 days at 120 rpm. Then, 5 ml of each isolate culture was centrifuged at 3000 rpm for 20 min. Phosphate concentration in culture supernatant was estimated as described by Olsen and Sommers71 . Three replicates were used, and a standard calibration curve was made with KH2PO4 solution (Sigma-Aldrich).
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