Electrophoresis Synthesis Lab Report

We use eight lanes putting the enzymes, and the DNA of the suspects to the lane. It is important to do this slowly and with care so as to ensure we put each solution in the correct tubes. You also have to plug it to the power source. Red is the negative while black is the positive and it should be at 150V for about forty minutes. Evaluation of the DNA bands

The idea behind Gel Electrophoresis is that we inject a slab of gel with the DNA we found at the crime scene. We then inject the same gel, next to the crime scene DNA, with suspect 1’s DNA and suspect 2’s DNA. We then send an electric current through the gel and wait for the results. The smaller molecules in the DNA will travel farther than the bigger molecules because the bigger ones will have difficulty making its way through the microscopic beads in the gel. After the separate bands appear in the gel, we stain it with a special chemical called Ethidium Bromide to give it a color under the blue

We placed the gel into the running chamber, and then we completely covered the gel with TAE. 3 microliters of loading dye was added to each tube; this would help distinguish the enzyme from the gel. As before, we tapped the tube on the table to mix. Then we carefully added each of the four samples into their own wells. A total of 33 microliters of each sample was poured into each well. Afterwards, we attached the positive and negative electrodes to their corresponding terminals on the power supply and gel box. We turned on the power to around 80 volts and waited 45-60 minutes for the loading dye to move down the gel approximately 6-8 cm. Finally, we were able to visualize the DNA in the gel and write down the

After electrophoresis was finished, my boss removed the gel casting tray from the running chamber. Then, she carefully transferred the gel to a DNA staining tray. My lab assistants and I were going to stain it so it would be easier to see the

To perform this event, it took approximately 0.4g of agarose per 50ml of solvent or 0.8g of agarose per

In slab gel electrophoresis agarose or polyacrylamide is used. The conducting buffer is contained in the porous gel and the gel is then poured in between glass plates. These plates are separated by spacers. An electric field is applied at the rear after the substance of experimentation is added to the wells at the top. The substance will then move a certain distance towards the positive electrode. This method uses gel unlike the cellulose acetate strips but it still uses a buffer and a electric field to migrate the substances based on their size and charge. Slab gel electrophoresis is usually used in the biological

Purpose The main purpose of this experiment was to test and observe how DNA molecules are being tested or separated. Introduction The final goal of this lab was to successfully measure the size of different samples of DNA placing each samples into a well in agarose gel and running a current through a charged chamber.

Gel electrophoresis is a gel technique that separates DNA and proteins based on their mass, by means of an applied electrical field that passes through an agarose or polyacrylamide gel. The concentration of agarose in the gel is commonly 0.8% to 1.0%, since agarose is expensive. The gel is embedded on a buffer, pH of 8.3, which keeps the pH of the solution at equilibrium. Assuming that at typical pH, DNA is negatively charged, denatured protein samples are placed in the wells located on the negative electrode side; hence the positive electrode side is located at the other end of the gel. As the positive and negative electrode sides are connected to a power source, protein sample migrates to the positive side of the gel. Migration of proteins is not necessarily based on their mass or mass to charge ratio; protein migration across the gel is based on their size. In other words smaller molecules will travels further than bigger molecules. Since the SDS gel contains agarose or polyacrilamide, molecules have to be small enough to migrate to the other end of the gel without getting stuck on the way.

The separation of macromolecules for laboratory analysis can involve the use of any of several techniques that isolate macromolecular components on the basis of size, solubility, reactivity, and volatility. Electrophoresis, for example, is a method of macromolecular separation that involves the use of a suspending medium, known as the matrix, and an applied electrical current that separates molecular components based on varying size and reactivity. While a uniform electrical current may activate the movement of molecular components through the matrix, the sponge-like structure of gel matrixes typically used in nucleic acid electrophoresis, such as agarose or polyacrylamide, inhibits the uniform migration of these components through the matrix

Dr. Park had created the gel for the class. I added 10 μl of the 5x PV92 XC loading dye into the PCR sample. I added 10 μl of the MMR-DNA marker into lane one, 10 μl of the homozygous (+/+) into lane 2, 10 UL of the homozygous (-/-) into lane 3, 10 μl of heterozygous (+/-) into lane 4, and 20 μl of my sample into lane 5. After the other students loaded their sample into the gel, I hooked up the electrodes and turned on the electrophoresis apparatus, which was at 100 volts. The electrophoresis apparatus ran for 40 minutes, which was until the marker dye had traveled half of the distance to the end of the gel. I then removed the gel and submerged it in the Fast Blast DNA stain (100x) using the staining tray. To create the staining gel, we used the formula M_1 V_1= M_2 V_2. The initial concentration of the gel was 500x, but we needed 100 ml of a concentration of 100x. I calculated that we needed 80ml of H_2 O and 20 ml of the dye. I stained the gel for 2 minutes. I rinsed the gel in warm water for 10 minutes. I compared my sample lane with the control lane and recorded my

I added 1 g of the agarose powder to 100 ml of TAE buffer in an Erlenmeyer flask. Using a magnetic stir bar, the agarose is heated until it is a completely clear solution. The agarose is poured into the chamber to make wells deep enough to hold 30 µl of solution. I inserted the comb in the chamber and waited for the gel to harden. To prepare for the samples, 10 µl of the XC loading buffer was added into the PCR tube. The professor demonstrated the techniques to load the gel with the DNA samples. After learning the techniques, I added my PCR sample in one the the wells. To run the gel, the power supply was set to 100V for approximately 30 minutes. To stain the gel, the gel was removed from the electrophoresis chamber and was placed into the FastBlast staining tray. The gel was placed in the staining tray for 2 minutes. Then the gel was rinsed for 10 seconds in Wash Tray 1. After rinsing for 10 seconds, the gel was washed for 5 minutes in Wash Tray 2. After agitating the gel for every minute in Wash Tray 2, the gel was washed for another 5 minutes in Wash Tray

Let the agarose powder sit in the buffer for a few minutes. The beaker is covered with plastic wrap an placed in the microwave. Microwave the solution slowly with boiling it. As soon as the solution starts to boil, take it out and carefully mix it with the hot gloves on and continue to heat the solution until it is completely clear. Once the solution is cooled, add 3µL of ethidium bromide stock to the solution and mix it by swirling it. The gel is poured into the prepared mould that is taped on the ends and eliminate any bubbles. Place the comb on the negatively (-) charged side. After the gel solidifies, remove the comb and tape and place the gel into the chamber. Gently pour TAE buffer over the gel as the gel should be completely covered by 2-3 mm of buffer. If air bubbles form, gently displace them with a disposable micropipette tip. Droplets of prepared loading buffer are placed on a piece of parafilm paper. The ladder will consists of only the buffer because it will work as a measuring device to compare the DNA samples. 10 µL from each of the DNA samples will be mixed with the loading buffer using the micropipette tip. The positive control will be consist of 10 µL of water and the droplet of loading buffer. The samples including the ladder and positive control are added to a separate well in the gel. Once all of the samples are loaded into their own

Analysis of DNA from practicals 1 and 2 using the technique of agarose gel electrophoresis and analysis of transfomed E. coli from practical 2 (part B)

Gel electrophoresis proving to give a very positive and forward moving impact in the science field, has many advantages. For one this scientific technique caters a clear link

Agarose gel electrophoresis is a method of gel electrophoresis used in biochemistry, molecular biology, genetics, and clinical chemistry to separate a mixed population of DNA or proteins in a matrix ofagarose. The proteins may be separated by charge and/or size