While, 5µl of purified PCR product, 2µl buffer R, 1µl BamHI, 1µl HindIII and 11µl distilled water were added into SOD gene tube. Next, 1 µl of 0.2mg/ml RNase was put inside both tubes and spinned by using microcentrifuge for a few seconds. Those tubes were then placed into 37ºC water bath and incubated for 2 hours. The tubes were deactivated by heat at 65ºC for 10 minutes and stored in -20ºC. Results Measurement of DNA concentration Equipment Optical
Plasmid and PEI reagent were prepared both 1:3 and 1:5 ratios and incubated during 15 minutes at room temperature. Then, these plasmid-PEI mixtures were added to culture and incubate 24 hours and 48 hours for each ratios. The table 1 shows all prepared assay. Then, 10 µl MTT
The apparatus was then closed and turned on to run at 100 volts. Electrophoresis ran for 30 minutes; separating the DNA according to size in the gel. Then the tray with the gel was removed and a stain sheet was placed on the gel for 15 minutes. After 15 minutes that gel was rinsed with water and then set in water. Later, 20 to 30 minutes, the gel was view over a light box to view how far the bands traveled.
The samples were filtered by vacuum over pre-weighed and wetted glass fiber filters (24 mm GF/C; Whatman; Kent, UK). The samples on the filters were rinsed with 5 ml stop solution to remove non-specific, cell surface binding of 3H-FLC. The filters with fungal balls were either allowed to dry for 24-48 hr or were baked in a drying oven for 15 minutes at 95° C. Each dried filter containing fungal balls was then re-weighed to obtain the dry mass of each fungal sample. The filters were finally transferred to 5 mL scintillation vials containing 3 ml of scintillation cocktail (Ecoscint XR, National Diagnostics, Atlanta GA).
Screw the liquid onto both test tubes to make sure that they are sealed. You now have to wait for approximately two days, in order to obtain satisfying results. Light the candle/put it on fire. Fill the third test tube with approximately two millimeters of Ethanol.
Experiment and Results Sample 100 ng/µl DNA was extracted from the cricket Acheta domesticus using the phenol-chloroform methods described in Davies et al., 2012 , dissolved in Tris-HCl-EDTA (TE) buffer and kept frozen at -20˚C. In initial tests, portions of the extracted DNA were suspended at the same DNA concentration as the control sample in solutions of magnesium chloride, magnesium sulfate, ammonium sulfate, lithium chloride, and nickel chloride. Each salt was mixed in three different concentrations, including 100 mM, 10 mM, and 0.1 mM. Then DNA in each salt concentration was incubated at different temperatures: 25˚C, 42˚C, 65˚C, and 95˚C, for fifteen minutes. The products were then loaded onto an agarose gel and allowed to run in
We added 1 ml of distilled water to test tubes labelled 2, 3,4 and 5. . To tube 1 we added 1ml of standard protein solution and recorded the concentration of the standard protein as 10mg/ml. To tube 2 we added 1ml of standard protein solution and shake it so that it can mix well. With a fresh pipette we removed 1ml from tube number 2 and added it to tube 3 then gently shake the tube. With another fresh pipette we removed 1ml from tube 3 and added it to tube 4, shake well.
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.
Yellow FeCl3 5% solution is added to the nanoparticle solution with 5% ratio of FeCl3 solution: nanoparticle solution (3 drops: 1 ml). The mixture is then stirred slowly for 30 minutes. The colored nanoparticles solution is then observed with the light microscope (Advanced, Indonesia) using OPTILAB application. Total essential oil
The sample was then incubated at 56°C to lyse the tissue. The sample was checked every fifteen minutes and vortexed between each checking for an hour and a half until the tissue was completely lysed. The tissue sample was then again vortexed. Next 200 microliters of buffer AL was added and
Transformation was successful in the plates where the bacteria consumed the pGLO plasmid. In the first plate that the bacterium was plated on it included the LB broth and of ampicillin antibiotic (amp), 2 colonies were present. The second plate of bacteria was grown with the presence of LB broth, ampicillin, arabinose sugar (ara), and 22 colonies were observed. But a green fluorescent glow of the colonies was only present in plate 2. Plates 3 and 4 were the control plates.
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. Each sample was loaded into the gel as well as 10 μL of DNA size markers (1kb ladder, New England Biolabs) into a separate lane. The gel was allowed to harden at room temperature and then electrophoresed at 100 volts for 75 minutes. Using a UV imager, a photo was taken of the resulting traveled DNA fragments in the gel.