The washing step were repeated. Using (95% v/v) alcohol wash off the iodine solution until the washing are pale violet. Immediately rinse with tab water. Using the Safranin solution (0.5% w/v), cover the smear for 1 minute. Use tab water to rinse off the stain.
Add choloroform and methanol of ratio 2:1 and of equal volume. Mix the mixture and left it for overnight for the process of evaporation. The white coloured sediment appeared that is the biosurfactant. Drying of biosurfactant Poured the sediment in the sterile petriplate of known weight. Put the plate in the Hot Air Oven at 100℃ for 30 minutes.
the supernatant was discarded and the pellet re-suspended in 400 µl of TE buffer (40 mM Tris-Base, 20 mM acetic acid, 1 mM EDTA, pH 8.0). 6 µl of 7.5 M ammonium acetate was added and the pervious step was repeated. 700 µl of 70 % ethanol was added and spun for 2 min at maximum speed. The supernatant was discarded and dried at 37°C, then finally DNA precipitate was re-suspended in 100 µl of TE
The supernatant was assayed for SOD activity by following the inhibition of epinephrine auto-oxidation. 0.5ml of sample was diluted with 0.5 ml of distilled water, to this 0.25 ml ethanol, 0.5 ml of chloroform (all reagents chilled) was added. The mixture was shaken for 1 min and centrifuged at 2000 rpm for 20 min. The enzymatic activity in supernatant was determined. To 0.05 ml of carbonate buffer (0.05 M, pH 10.2) and 0.5 ml of EDTA (0.49 M) was added.
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.
The graphite rod was inserted as anode into the ionic liquid (IL)/water solution, placed parallel to the Pt wire as counter-electrode with a separation of 1 cm and the ionic liquid urea choline chloride was mixed with water at 1:1 ratio. Using a DC power supply the potentials of 5 V were applied to the two electrodes. The exfoliation products were washed with water and surfactant until the pH became neutral and the products were separated by ﬁlter and ultra-centrifugation at 10,000 rmp at 25 ºC2. 2.2 Preparation of SRGO Synthesis of SRGO was carried out using the method described by He et al.24 and the process of SRGO production is shown in Fig. 1.
Then the saline was expelled back into the cup. 1,000μl of the oral rinse was transferred into the 1.5μl micro centrifuge tube. The tube was then spanned in a balanced centrifuge for 5 mins in order to make a pellet of whitish cells at the bottom of the tube. The supernatant was discarded and the pellet was re suspended by the process of vortexing. After this process 20μl of the re suspended cells were added into a tube containing Instagene Matrix.
Not enough time was given for the precipitate to settle in the beaker, hence when the supernatant is removed; there is a large amount of product lost. This is especially so in step 25, because a large amount of solvent is decanted. It will require much longer than the given time of 10 minutes to allow most of the precipitate to settle at the base of the beaker. Hence instead of decanting the mixture, centrifugation can be used to remove the solution instead. The amount of product lost will be significantly lower by carrying out this process.
The reaction mixture was poured into ice cold water acidify with 1% HCl and precipitates were collected, filtered and dried and recrystalized with ethanol. 4.2.2 General procedure for synthesis of flavones: 50mg of chalcone was taken in round bottom flask. After that 15-20 ml of DMSO was added and mixed properly, 5-6 granules of iodine were added and reflux the reaction mixture up to 3-6 hours and kept for overnight. Then reaction mixture was poured into ice water and the precipitates were neutralized with sodium thiosulphate, to remove the unreacted iodine, washed with water, filtered, dried and recrystalized with