Cool the filtrate to room temperature, add 1.5 g of sodium nitrite, and stir until reaction is complete. 5. Pour this mixture, while stirring, into a beaker containing 25 mL of ice water to which 5 mL of concentrated hydrochloric acid have been added. The diazonium salt of sulfanilic acid should soon separate as a finely divided white precipitate. Keep this suspension cooled in an ice bath until it is to be used.
Heat the Agarose gel in a 65 °C water-bath to melt the agarose. After it melted, maintain its temperature at 55 °C until it is ready for use. 2. Transfer the spheroids from the 96-well plate to a 15 mL centrifugal tube using a 1000 μL pipette 3. centrifuge the tube for 5 min at 1000 rpm to form a pellet. 4.
After the finalization of alkali formulation to be used for extraction of cornhusk fibres, the fibres were treated with Pulpzyme HC to increase their fineness. Pulpzyme HC is a xylanase enzyme (EC.3.2.1.8) produced by submerged fermentation of a genetically modified Bacillus microorganism, and was obtained from Novozymes. The enzyme has an activity of 1000 AXU per gram. (http://fzfz.nbdl.gov.cn:81/files/20130815/1376527490502_36.pdf) Effect of concentration of Pulpzyme HC and Treatment Time The extracted fibres were treated with three different concentrations of enzyme i.e. 1%, 3%, 5% (owf).The time taken for treatment was 30 minutes, 60 minutes and 90 minutes.
22.5 g of plate count agar powder was dissolved in a litre of sterile distilled water on the hot plate 2. pH of the solution was adjusted to 7.0 ± 0.2 by adding NaOH or HCl and was immediately transferred into the Schott bottle to be autoclaved at 121 ° C for 15 minutes 3. Prepared medium was stored in 4° C chiller Lauryl Sulphate Broth
It is to prevent the cell from washing away during the staining and washing process. Then, it is air dried and followed by fixing it with flame from Bunsen burner. After fixing the smear, it must be stained using Gram staining solution, firstly crystal violet solution was flood onto it, and allowed for 1 minute, then wash off with tap water. Then, flood the slide with iodine solution for 1 minute and wash it off with tap water again. The formation of a dye-iodine complex will occur in the cytoplasm.
2.2.Yeast cell preconditioning and inoculum preparation 1 g dry weight of yeast was resuspended in 100 mL of deionized water in an Erlenmeyer flask of 250 mL volume, at 30–35°C, for 30 min with NaCl 6% w/v. Inoculum for experimental fermentations was prepared
Pour 50 ml of 1% agarose solution kept at 50ºC onto the casting plate. Immediately place the supplied comb about 1 cm from one end of the plate ensuring that teeth of the comb do not touch the glass plate. Wait till a firm layer of gel is
H2SO4 • Catalyst: potassium sulphate (K2SO4), copper (II) sulphate pentahydrate (CuSO4.5H2O) • NaOH (40%) • 0.1 N HCl solution • Boric acid (4%) • Indicator: methyl red (200 mg make up to 100 ml with %95 ethanol ) 1 gram Feta Cheese was used as a sample in this experiment. First of all, the experiment was started with the digestion phase which was performed by mixing 10g potassium sulphate and 0.5 g CuSO4 solution that acts as catalyst, and added 20ml conc. H2SO4. The digestion unit temperature was set to 420±10°C and preheated for 15 minutes. Then, it was heated continuously at least 2 hours till a clear colorless solution was obtained.
Using distilled water would be a good method in order to rinse the beaker. Make the solution up to the 500cm3 mark with iodine (1% concentration) • Starch Indicator Solution Weigh 0.25g of soluble starch and add it to 50cm3 of near boiling water in a 50cm3 beaker. Stir it in order to dissolve and wait for it to cool before using. Procedure Safety • Before conducting the experiment, be sure to put on safety goggles and
200ml of water was then added to the filtrate in a 500ml beaker with constant stirring. White solid was formed in the process of addition and the solution was then left undisturbed in an ice bath for 10minutes. Once most of the solids had settled at the base of the beaker, the solution was decanted. 10ml of ethanol was added to the remaining suspension and was transferred in a clean centrifuge and centrifuged for 2minutes at 6000rpm. After the first centrifugation, the supernatant was discarded and the residue was washed by adding 14ml of ethanol.