Then, water was added dropwise during the mixing process. The above solution converts of colorless to yellow suspension solution which produced TiO2 nanopowder by drying process at 85°C in anstove for 15 hours. Finally, TiO2 nanopowder obtained were treated in furnace at different temperatures (400°C-800°C) for 2 hours. The initial heating rate was maintained at 5 °C/min. 2.3.
In 10 g dried sediment sample added 7 ml 0.2 M NH4Cl solution. A mixture of 100 ml hexane: acetone (1:1) was used as a solvent to extract pesticides with overnight shaking for 12 h on reciprocal or wrist action shaker at 180 rpm. The extract was carefully decanted through activated florisil column (2-3 cm), giving twice wash with25 ml hexane: acetone (1:1) to the sediments. The elute was then washed with 200 ml water and then again aqueous layer was extracted with 50 ml hexane. Finally the hexane layer was washed with 100 ml water and then evaporated to dryness with a vacuum rotary evaporator.
The incubation mixture contained 2.5 ml of 1.2% (w/v) fibrin, 2.5 ml of 100 mM Tris–HCl buffer, 10 mM CaCl2 (pH 7.8), and 20 µg of enzyme. The incubation was carried out at 37°C for 30 min, and the reaction was stopped by adding 5 ml of 110 mM trichloroacetic acid containing 220 mM sodium acetate and 330 mM acetic acid. This reaction mixture was centrifuged at 3,000×g for 5 min, and the absorbance of the trichloroacetic acid (50 mM) soluble product was determined at 275 nm. One unit of fibrinolytic enzyme activity was defined as the amount of enzyme required to liberate 1 µg of L-tyrosine per minute at 37°C. The total protein determination was performed as described by Lowry et al.
In the round-bottom flask (100 mL), we placed p-aminobenzoic acid (1.2 g) and ethanol (12 mL). We swirled the mixture until the solid dissolved completely. We used Pasteur pipet to add concentrated sulfuric acid (1.0 mL) to the flask. We added boiling stone and assembled the reflux. Then, we did reflux for 75 minutes.
Apparatus- Chromatography column: C18 (10 microns particle size), with Guard column Flow rate: 1.2ml/min Pressure: 30-40kgf Wavelength: 326nm Mobile phase: methanol : water (95:5 v/v) Internal standard: retinyl acetate Injection volume: 20µl Procedure for Retinol extraction from serum samples- 1) 100 µl of serum sample and 100 µl of Retinyl acetate were added into 12 X 100mm glass test tubes. Vortex-mixed for 30 seconds. Then, kept them at 4 C for 5 mins. 2) 1mL of hexane was added and vortex-mixed intermittently for 60 sec. 3) Centrifuged at 2500 rpm for 12 mins.
Then the blower was turned on for sufficient duration and UV lights were switched on for one hour. LAF microbial contamination test (Settling plate method) A total of three petri dishes were prepared aseptically inside a laminar air flow (LAF), and then the petri dish was filled by pouring sterile Tryptone Soy Agar (TSA) liquid media and allowed to solidify. One test media was placed in the LAF cabinet, one test media is placed beside the LAF cabinet while the other the test media was placed near the door. The three petri dishes lid were opened and allowed to stand for 15 minutes and closed again. Then, the test media is then incubated at 37 ° C, for 18-24 hours.
• Hydrogen peroxide (H2O2, 2mM) in phosphate buffer (3.0ml) was taken in an experimental cuvette, followed by the rapid addition of 40μl of enzyme extract and mixed thoroughly. • The time required for a decrease in absorbance by 0.05 units was recorded at 240nm in a spectrophotometer (Genesys 10-S, USA). • The enzyme solution containing H2O2-free phosphate buffer
The cooling curve was determined by recording the temperature at regular time intervals (every fifteen seconds) as the cyclohexane cooled, until the temperature became constant. The cyclohexane was stirred continuously. The FP tube was then removed from the ice and the cyclohexane melted. The cooling of the cyclohexane was then repeated twice and the mean of the three values was taken as the freezing point. ~0.1g of benzoic acid was accurately weighed and added to the cyclohexane and stirred vigorously until it had dissolved.
After adding three boiling chips, 10 mL of 48% hydrobromic acid was also added to the round bottom flask and swirled for 15 seconds to reactants in the flask. The reactants were clamped to a ring stand and a pre-set reflux apparatus with clear hoses attached to the condenser. The voltage regulator was set to 40 to begin water flow through the condenser and the application of heat, so the solvent can boil. The reaction was set to reflux for 30 minutes. Upon completion, the round bottom flask cooled for three minutes in a beaker filled with room temperature water and again in a beaker with ice cold
0,1 gr of kaffir lime oil nanocapsule is weighed and diluted to 100 ml using aquadest, taken as much as 1 ml (100x dilution) to put in the reaction tube then 1ml of saturated NaCO3 solution is added to the test tube and incubated for 10 min at room temperature. Then 0.5 ml of the folinciocalteu (Chemix CV, Yogyakarta) reagent and 7.5 ml aquadest were added, the mixture homogenized using vortex and then incubated for 30 min at room temperature under dark environmental conditions. Absorbance of the sample was then measured using a UV-vis spectrophotometer at a wavelength of 770 nm. The total phenol content of the sample was interpreted to be equivalent to gallic acid based on the standard curve of obtained gallic
One sample had 250ng of plasmid A as well but with no enzymes added. All the digestions tubes were incubated at 37℃ for 30 minutes. After incubation, 5μL of loading buffer (30% glycerol, 10 mM Tris-HCl, pH 8, 1 mM EDTA, 0.025% bromophenol blue) was added to each sample. 50 ml of molten agarose (1% agarose boiled and cooled to 55℃ with added SYBRsafe) was poured into the casting tray for gel electrophoresis. Once the gel hardened, .5X TBE (44.5 mM Tris base, 44.5 mM boric acid, and 1.0 mM EDTA) was added just until the gel was covered with the TBE buffer.