The solvents DMF and methanol were distilled for purification. Other chemicals were used as obtained. 2.2 Preparation of polystyrene (PS) Polystyrene prepared by free radical polymerization of styrene monomer. Styrene (1 mole) was taken in a round bottom flask (RBF) fitted with a reflux condenser. DMF was used as a solvent and AIBN (0.5% w/w of total monomer) as free radical initiator .The reaction was carried out at 70±2° C for 6 hour with constant stirring.
The tube was placed back in incubation for 96 more hours to observe any more positives. 2.10 Catalase Test A trypticase soy agar plate was used and after incubation, four drops of 3% Hydrogen Peroxide was added to the plate to flow over the bacterial growth. A presence of bubbling was observed. 2.11 Starch Hydrolysis A starch agar plate was inoculated with a streak of the unknown bacteria and then incubated. On the second day of incubation, the plate was removed from the incubator and placed over a hot plate heating Iodine solids.
To the double digest tube was added 12 μL double digest master mix, and 1.9 μL sterile dH2O. To the single digest tube was added 11 μL single digest master mix, and 2.9 μL sterile dH2O. To the undigested tube was added 10 μL undigested master mix, and 3.9 μL sterile dH2O. Digests were then incubated at 37 °C for one hour and then stored at -20 °C for one week. After which the digestions were examined by gel electrophoresis.
After a gram stain was done unknown #257 was identified as a gram positive organism because when observed under the microscope the organism appeared purple with cocci in clusters. The organism was also catalase positive which means that it produced enzyme catalase and bubbled when hydrogen peroxide was added to it. Three test were conducted based on the result of the gram staining procedure. Blood agar with a Novobiocin disk was chosen as well as DNase (DNA) and Mannitol Salts (MSA) agar. The Blood agar is a bright red, opaque plate and the streaking or the inoculation technique was a modified streaking for isolation with a heavy quadrant one.
10- Transfer the ester layer to a small dry test tube and dry the ester with anhydrous CaCl2 and stir for 10 min. 11- put it in a preweighed dry round bottom flask . 14- Determine the yield, refractive index, and % yield of ester. Conditions :- 1) This reaction is catalyzed by acid, Like Fischer esterification. 2) Usage of water in step (5):So that after Estrification is completed , any excess unreacted acetic anhydride is hydrolyzed.
The first of the four test was a Gram-stain. Once the experiment was completed the slide was then placed under a microscope where it was then categorized as Gram-negative bacilli. Its pink color when viewed through the microscope indicated that the test was Gram-negative and its rod shape indicated the morphology was bacilli. With these results the next test to be conducted was the citrate utilization test. A citrate tube was inoculated with P. aeruginosa and incubated at 37 degrees Celsius at 24 hours.
Rose Bengal-(bis(aminoethyl)ethylene glycol) (2) from Rose Bengal disodium salt (1) The synthesis was done following procedure from . Rose Bengal Na+ salt (915 mg, 0.90 mmol) was dissolved in DMF (2ml) and DIPEA (0.312 ml, 1.80 mmol), HATU (308 mg, 0.81 mmol) were added. After activation for 15 min, the mixture was added to O-Bis-(aminoethyl)ethylene glycol trityl resin (309 mg, 0.31 mmol) preswollen in DMF for 2 hours. The coupling reaction wrapped in aluminum foil was allowed to proceed overnight on a nitrogen bubbler apparatus. The resulting red-burgundy coloured resin was filtered and washed well with DMF.
The solution homogeneity expelled, by centrifugation for 10 min. The sample was centrifuged and separated into two layers, and took the top of the sample is injected for HPLC (11,12). Measured concentration in total lipid: The total fats balanced concentration of the pesticide getting by dividing the measured pesticide residue concentration in the overalll tissue sample by the decimal fraction of the sample that consisted of ether-extractable lipid. The total lipid content of each specimen was estimated from its total cholesterol & triglycerides levels by using a summation method. Analytical results for organochlorine pesticides were reported on a lipid-adjusted basis (nanograms per gram or parts per billion) (14).
Once the cola starts to boil, continue to boil it for another 10 minutes so that the carbon dioxide is removed. When the cola has finished boiling, cool it in an ice bath and pour the cola back in the volumetric flask and use distilled water to fill the flask to compensate for the evaporated water. Using a volumetric pipette, transfer 60ml of the cola to a beaker and put the magnetic stirrer in the beaker. Submerge the conductivity probe in the cola. Fill up the burette with NaOH
3.3.4 Preparation of CaCl2 solution The molecular weight of CaCl2 was 111 g/mole. Weighed 11.1g CaCl2 and dissolved into 100ml distilled water. The final CaCl2 concentration would be 1M. CaCl2 solution was used to make bottom agar and initiate the infection cycle. 3.4 Selection of most sensitive strain to bacteriophages to make new stock culture Starting culture was prepared by inoculating 1ml (1×109 CFU/ml) stock Lactococcus lactis ssp.