When the gel was polymerized, isopropanol was removed on the top of the running gel with water and thus, water was also removed as well. Then, stacking gel was prepared and its polymerization reaction was started with TEMED, and its solution was quickly poured on the top of the running gel. Therefore, a comb was inserted into the stacking gel before its polymerization, to form the loading wells. When the gel was polymerized, the inner chamber was filled with electrophoresis running buffer and the outer chamber was filled half. Chemical modification of ERAB (Wild Type and Mutant) Protein with Fluorescein Maleimide Wild type and mutant ERAB
Next, I dye the Unknown with Gram’s iodine to create a complex only have on gram positive. The slide is rinsed by water after 30 seconds. Decolorization is the next step of the whole process. I let the alcohol flow on 45-degree angle slide within 15 seconds and wash it with water to remove colors on the surface. Lastly, the unknown is once again dyed with safranin for 1 minute then wash it off with water for the last time and dry it using bibulous paper.
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
We then weighed a filter paper which we will use later in the experiment. We then collected the crystallized acetylsalicylic acid by vacuum filtration in a Buchner funnel and washed the product with a little ice-cold water. We then pre-weighed a clean, empty watch glass and labelled it with our initials and the date, we did this do we could easily identify that it was ours when we go to weigh it with the crystals on. We
Antimicrobial activity The agar disc diffusion method was employed for the determination of antimicrobial activity of the papaya leaf extract stabilized gold nanoparticles. The 0.1 ml of 108cfu/ml of different pathogenic bacteria suspension was spread on different plates nourished with LB media. Filter paper
Gather materials (beaker, 100 milliliters of copper chloride, graduated cylinder, funnel, 2 centimeter by 15 centimeter aluminum foil strip, goggles, gloves, apron, hydrochloric acid, and a spoon to push down the aluminum foil so that it doesn’t rise up and drip copper chloride down the side) 2. Pour 100 milliliters of copper chloride into the graduated cylinder to measure it 3. Pour the copper chloride into the beaker or leave it in the graduated cylinder (whichever you want to conduct the experiment in) 4. Fold the tin foil in half the long way so that you have 1 centimeter by 15 centimeter strip of aluminum foil 5. Curl it into a coil/spring 6.
Shake funnel and rinse off water layer ( This contains the sulfuric acid and majority of methanol). Again wash the ether with 25ml of water and then wash the organic layer with 25ml of 10% sodium bicarbonate to extract unreacted benzoic acid. Again shake separatory funnel with frequent venting of pressure and opening the stopcock. Allow the separation of layers and drain off bicarbonate layer into a beaker. Wash ether layer with saturated sodium chloride solution and retain ether layer.
4 Procedure Preparation of Test Solution The sample was prepared by adding 100 mg of powdered in 10 mL of methanol, soaked for 15 min. Centrifugation was done and the filtrates or supernatants were used as the sample solution. Out of this filtrate 50 μL of samples were dissolved in 1 mL of toluene and were used as the sample solution.
It is essential to use the leaves when they are as fresh as possible, do not leave them unattended after the measurements are taken. Step 6: Place the crushed leaves into the bottom of the glass jar of isopropyl alcohol so that they cover the bottom. Step 7: Place the lid on the glass jar tightly. Step 8: Pour hot water into a large bowl and place the glass jar in it. Step 9: Leave the glass jar in the hot water for an hour and shake the jar gently sideways from time to time to stir the leaves.
The key enzyme in the process is DNA polymerase that forms the PCR product by linking individual nucleotide bases (adenine, guanine, cytosine and thymine). Short DNA fragments with sequence complimentary to target DNA are called primers; these specify the DNA sequence that has to be amplified. The PCR reaction mixture is made in a 96 well plate or test tube and placed in a thermal cycler. The thermal cycler is a machine used to amplify DNA by altering the temperature in repeated cycles using three precise pre programmed steps: Denaturation – By increasing the temperature in the thermal cycler the DNA is denatured creating separate single stranded DNA
This procedure occurred in the presence a Bunsen burner. The inoculation needle were placed within the open flame 15-20 second in order to sterilize the needle and prevent contamination. The needle was allowed to cool 5-10 second before inoculation. Using aseptic transfer technique the needle was used to gather up some of the colonies on the plate, making sure not to touch anything else with the needle. The test tube was uncapped and the lips of the test tube was passed through the open flame three times.
The Solid sequencing platform, produced by Technologies/Applied Biosystems (ABI), performs sequencing by ligation method. Similar like the Roche 454 library preparation, genomic double strand DNA were sheared into small pieces and ligated with two types of adatptors P1 and P2 on two ends. One end with P1 adaptor binds onto the surface of the magnetic bead and emulsion PCR takes place to amplify single nucleotide fragment. Then the oil was washed out and four fluorescent labeled di-bases probes were added into the beads mixture. By matching the 1st and 2nd position of the template by di-base probes, fluorescence was detected and the extra tail with fluorescent probe is cleaved out.
The purpose of this experiment is to create a complete genomic library of Aliivibrio fisheri through the use of the lux operon. The examination of the lux operon gene occurs through the extraction of the DNA of Aliivibrio fischeri and digest a large piece of DNA to smaller random pieces. The fragment of DNA will later be ligated together in plasmid. Plasmid acts as vectors to transport DNA from one organism to another. The DNA will then run through a UV-visible spectrophotometer to test the absorbance of the extracted DNA.
Next one microliter of DNA was mixed with one microliter of loading dye using a pipettor and loaded into the well. The same mixing process was completed for the PCR product, using one microliter of PRC product and one microliter of loading dye. For the purposes of this experiment, the DNA product was loaded into well six and the PCR product was loaded into well seven. Initially DNA was loaded into well five, however gel was pierced so samples were moved one well to the right. The gel was run at 100 V for one hour.