17) The outcome was cataloged. 18) The left and the right beaker are emptied and cleaned to execute the next test. 19) 18 millimolar of Na⁺Cl⁻ was put inside of the left beaker. 20) Deionized Water was poured into the right beaker. 21) A sixty-minute timer was started to see how the 18 Na⁺Cl⁻ (mM) solvent diffuses through the 50 (MWCO) Dialysis Membrane.
A control extract is prepared (5ml of DAE) to a test tube, which is then placed in boiling waterbath for 10minutes, after 10minutes remove the control extract and leave it to cool at room temperature. In order to determine the amylase activity, one drop of iodine is dropped into 21 labelled wells on the ceramic test plates. A reaction mixture is prepared, 5ml of buffer and 1ml of 0.5% starch solution to a test tube. Extract one drop from the reaction mixture to the well labelled T. Turning blue-black indicating the presence of starch in the reaction mixture. Add 1 ml of diluted amylase extract to the reaction mixture.
Sample Preparations Unknown water and reagent blanks are prepared in the same manner. 20 ml test tubes are taken and 5ml of water is pipetted into each of the test tubes. The pesticide standards are now inserted into the test tubes at concentrations of 0.1,0.2,1,2,5,10 and 30 ng/ml-1. The mixtures are mixed and kept in equilibrium for 30 minutes. After the 30 minutes is done, the tests tubes are then immersed in a 100 degrees Celsius methanol water bath for 15 minutes.
The acid was poured into the flask until there was a permanent pink colour. The acid was allowed to be poured for a little longer before the flask was removed and taken to a lab bench with a buret that contained 0.1 M sodium hydroxide, and the amount of acid used was recorded. The sodium hydroxide was added into the flask in small amounts
Standard Preparation: 100 mg of standard ascorbic acid was weighed precisely and transferred to a 100 ml volumetric flask, added 70 ml of 0.5% sodium metabisulphite and dissolved by shaking. The volume was made up to the mark with 0.5% sodium metabisulphite for getting a concentration of 1 mg/ml. 2 ml of this solution was taken into another 100 ml volumetric flask and made the volume up to the mark with 0.5% sodium metabisulphite which resulted in concentration of 0.02 mg/ml. The solution was filtered through 0.45 µ nylon syringe filter. Sample Preparation: 2.5 g of sample was weighed accurately and transferred to a 100 ml volumetric flask.
This diluted solution will be used in the assay as duplicate samples. Then, 1.0mL of standard glycine solution containing (7.5mg/mL) was diluted to 100mL with water using a volumetric flask. This solution contains 1.0µmole/mL of glycine. 8 tubes were set up according to the following protocol and 2.0mL of ninhydrin reagent was then added to each of the 8 tubes and were placed in a boiling water bath for 20 minutes. After 20 minutes, the tubes from the bath was carefully removed, cooled in a beaker of cold water, then 8.0mL of 50% ethanol was added and mixed well.
After which the digestions were examined by gel electrophoresis. The samples were run on a 50 mL 0.9% (w/v) agarose gel in 1X TAE buffer at 100 V until the leading track dye traveled 2/3 the distance of the gel. The gel was then soaked in GelRed for 20 minutes and examined under UV light. To prepare the digestions 10 μL of each digestion was mixed with 2 μL of 6X track dye in a micro centrifuge tube. 12 μL of 1 kb DNA ladder and each digestion was run on the
Subsequently extracted by Microwave at power level 70 for 16 minutes and then filtered. The filtrate obtained, was added 10% HCl (until pH 2-3). Then do the bleaching with NaOCl diluted with water 1: 1 to white. Then converted to sodium alginate by adding 20 g of Na2CO3 and stirred in a mixer. The resulting solution is then etched with ethanol to form sodium alginate fibers.
Absorbance versus Time Measurements: The absorbance was set to 0 Abs while the spectrometer was set to ʎmax (from Part A). In Part B, 1.00 ml of the solution was mixed with the Blue dye in the beaker and half-way covered with a cuvette. Concurrently, the Spectronic 20 was blanked with water. The processes detailed above were repeated, each at a time. The absorbance values were recorded for 11 minutes.
Every 20 min, 30 ml of sterile distilled water was add¬ed into the outside buffer and repeated 5 times to make a dilutional factor of 2.5. The dialysis bag was transferred into 100 ml of isoos¬motic buffer; pH 7.45 and incubated at 37°C for 1 h. Generated pores of RBCs’ membrane loaded with FVIII reversed into normal in isotonic media. The isoosmotic buffer was contained: 0.036 mM KH2PO4/KOH, 0.04 mM MgCl2, 0.84 mM NaCl, 0.18 mM glucose, and 0.27 mM adenosine. The RBC-carriers washed 3–4 times in sterile phys-iological saline by centrifugation (1250 g, 10 min) to remove hemoglobin and unloaded factor