Figure 20 and 21 casting tray Figure 22 gel box After 1 hr., take out the gel from gel box carefully, place it into the machine, so that the DNA gel electrophoresis can be visualize under UV light. Figure 23 gel documentation system, used to visualize gel electrophoresis with UV light NMR spectroscopy First of all, 3 samples were prepared, peptide in SDS, DPC, and buffer. The sample temperature was maintained at 298 K, prepared by supervisor and H(hydrogen) in SDS and DPC micelle was replaced with D(deuterium) , so that in proton NMR, peptide won’t be interfered by H in micelle. amount of peptide in sds and dpc In the case of NMR of peptide alone, sample was prepared with 90% of H2O which is 540μL and 10% of D2O which is 60μL, so 600μL of solution was used to dilute 1.5mg of peptide. Put 600μL of sample into NMR Sample Tubes, put the sample tube into NMR sample holder, and then run the test, the chemical shift of proton in peptide can be monitored.
PCR product (5 µl) was added to each well and loading dye (5 µl) was added to the final well. The PCR chamber was then sealed and a current of 110mA applied for one hour. The solidified gel was then removed from the chamber and placed under UV light. A digital image was then taken, allowing for analysis to be carried
The incubation mixture contained 2.5 ml of 1.2% (w/v) fibrin, 2.5 ml of 100 mM Tris–HCl buffer, 10 mM CaCl2 (pH 7.8), and 20 µg of enzyme. The incubation was carried out at 37°C for 30 min, and the reaction was stopped by adding 5 ml of 110 mM trichloroacetic acid containing 220 mM sodium acetate and 330 mM acetic acid. This reaction mixture was centrifuged at 3,000×g for 5 min, and the absorbance of the trichloroacetic acid (50 mM) soluble product was determined at 275 nm. One unit of fibrinolytic enzyme activity was defined as the amount of enzyme required to liberate 1 µg of L-tyrosine per minute at 37°C. The total protein determination was performed as described by Lowry et al.
The swab was inoculated onto blood agar, MacConkey’s agar and nutrient agar, mannitol salt agar and Sabouraud's dextrose agar and was incubated at 370C for 24 hrs for bacterial culture and at room temperature for fungal isolates. Next day the colonies were picked up and preliminary identification was done and the bacterial isolates were identified based on standard protocols. LPCB was done for the identification of fungal isolates after 48 to 72 hrs. ANTIBIOTIC SUSCEPTABILITY TEST Antibiotic sensitivity test of the bacterial isolates was determined by the Kirby-Bauer disc diffusion method2. The following were the antibiotics used for the study-amikacin (AK 30), nalidic acid (NA 30), erythromycin (E 15), vancomycin (VA 30), tetracycline (TE 30), cefoxitin (CX 30), rifampicin (RIF 5) ciproflaxicine (Cip 5), ceftazidime (CAZ 30), cefotaxime (CTX 30), cepifime (Cpm 30), cefoperazone (CPZ 75).
The effect of DMBQ against S. typhimurium cells was bacteriostatic for the first 5 h of incubation after addition of the compound. The bactericidal effect DMBQ was observed against S. aureus after 12 h of incubation. Thus, doubling the MIC of DMBQ reduced the growth rate of S. aureus but did not have a siginificant effect on the final cell
The solution was stirred at room temperature for 8h. The solvent was blown out with nitrogen. The residue was added to 1 ml of water containing 0.1% TFA and purified on RP-HPLC. Massspec of the final product clearly indicates presence of RB modified on PEI by series of peaks matching different polymer compositions (see Fig. 6).
An equivalent volume of saline solution was added (NaCl 0.16 M; pH = 7) and it was mixed completely. The resultant solution was centrifugated for 30 minutes at 9000 rpm at 4ºC in a rotor Beckman JA-20, eliminating the pellet. The supernatant was collected to obtain the lipoproteins by gradient density ultracentrifugation: 10ml of the yolk solution were taken and then 0.9 g of potassium bromide (KBr) was added. The bromide was dissolved with a smooth agitation, with care not to denature the proteins. Straight afterwards, saline solution was added (NaCl 0.16 M, pH=7) to the 10ml of the yolk solution with a Pasteur pipette, avoiding the sample diffusion, forming two phases and filling the tube completely.
The bacilli can be carried by droplet infection. DROPLET INFECTION : With the process of coughing and sneezing, droplets may be ejected right onto the face. Bacilli thus intiated easily take up their place in host without losing much of its vitality. Human type can flourish best when they are immediately carried from one mass to another. They begin to lose their vitality outside human body although they are known to cling to dust, screen, clothes, utensils of the room for several weeks.
10. Now place the gel along with UV-transparent sheet on a UV-transillumintor and view the gel in UV light for presence of orange colored bands. 11. Measured the distance moved by each band from edge of the loading well. Draw a graph of log10 Mw of standard DNA marker vs the distance travelled by each of them 12.