The solution was discarded into the waste bin, and the materials were washed. The second reaction in Part B, sodium hydroxide and ammonium chloride, began by saving the data from the first reaction and setting up the LabQuest to new data collection under the same conditions as the first reaction. The cups were restacked and placed in the beaker. Using a graduated cylinder, 50mL 2M NaOH was added to the cup. The cup was then covered and the temperature probe inserted.
15 mL of Solution A and B were mixed together to form solution F. Eight cuvettes were labeled distinctly as 1a, 2a, 3a, 4a, 1b, 2b, 3b, 4b, where “a” cuvettes were used for the concentration experiment and “b” cuvettes were used for the temperature experiment. Cuvette 1, the blank tube was prepared and the spectrophotometer was set to 405 nm. The enzyme was added, upon being ready to start the experiment, to tube 1 which then became tube “1a.” 3 mL of solution F was added to each cuvette, both “a” and “b.” The “b” cuvettes were then placed in their specific temperatures, 1b in the fridge, 2b in room temperature, 3b in a 32 degrees Celsius water bath and 4b in a 60 degree water bath. The temperature was recorded using a thermometer that was placed in the surroundings of the tube. The cuvettes were retrieved from their respected conditions.
Measure rate of CO2 produced in the upside down 250 graduated cylinder (you will see that the water levels begin to decrease from the top of the cylinder. The gas at the top of the cylinder is CO2) 14. One individual will be watching the increase of CO2 and controlling the timer, while the other will be holding the test tube and recording the increase of CO2 15. At every 15 second interval, record the volume of CO2 gas produced in the cylinder, into a table, until you have reached 180 seconds (you must perform three trials for each concentration of HCL) 16. Carefully pour out all of the HCL that was in the conical flask and thoroughly rinse CaCO3 chips and conical flask with tap water 17.
Once this point was reached, the timer was stopped and the time was recorded in Data Table #1. The same steps involving the addition of Na2S2O3 were repeated for Wells #2 and #3, using 2 mL of Na2S2O3 in each. The final times for each well was recorded in Data Table #1 in the appropriate blank. Once the first trial was completed for the first three experiments, a second trial was completed in Wells #4, #5, and #6 using the same procedure in order to increase accuracy. After each trial was conducted, each of the wells were emptied into the designated waste container and dried using a paper
Finally the hexane layer was washed with 100 ml water and then evaporated to dryness with a vacuum rotary evaporator. The concentrated extract was then passed through a chromatographic column (30 cm x 10 mm i.d) containing 2 g florisil (lower) and 1 g sodium sulphate (upper) which is pre wetted with hexane: acetone (1:1). OCPs were eluted with 25 ml hexane: acetone (1:1).The solvent was evaporated using rotary evaporator and final volume was adjusted to 5 ml, which is used for GC analysis. All the sediments were analyzed for HCH and
• 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
13) Set the spectrometer to a wavelength of 530 nanometers. 14) Place the cuvettes (numbers 1-6) with the appropriate substance and record it’s reading in the data table. 15) After each cuvette was tested, place the distilled water sample (Cuvette zero) to reset the spectrometer and to ensure that the scale is calibrated and repeat for each cuvette test. Data/Results: Tube Number Concentration Of CoCL2 (Mg/ML) CoCL2 Stock (ML) Distilled Water (ML) Spectrometry Reading at
To this, 80ml of cold water and 15ml of 2M HCl was added to the conical flask. Afterwards, 0.1ml of ferroin solution (as an indicator) was added. Next, titration was performed. The contents in the conical flask was titrated with 0.1M ammonia cerium (IV) sulphate until a yellow solution was produced. The experiment was then repeated without sample B (only the H2SO4 and water in the proportion 3:7, 6ml acid 14ml
Place 20 MWCO membrane into membrane holder Step 3. Place sodium chloride with a concentration of 9.00 mM into left side of beaker Step 4. Place deionized water into right side of beaker Step 5. Run the experiment, once the timer hits zero record your data and flush all of the contents. Step 6.
for the determination of the minimum inhibitory concentration (MIC) of the active extract. The sStarting solutions of the tested extract were obtained by dissolving it in 5% dimethyl-sulphoxide. STwo fold serial dilutions twofold dilutions of the extract were made within a concentration range from 0.04 to 40 mg/mL in sterile 96-well plates containing Mueller–Hinton broth for bacterial cultures and a Sabouraud Dextrose SD broth for fungal cultures. Resazurin solution was added as an indicator to each well. and finally, to each well fungal or bacterial suspension was added .
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.
1. 150 ml of boiled water was poured into each of the three beakers labeled A, B, C. 2. Five tea bags were soaked for the time given by the manufacturer (two minutes) , in beaker A (Control). The teabags were immediately removed after the time elapsed. 3.