Dna Isolation Lab Report

1. If each individual has such a small amount of DNA, how do the bands on the gel contain enough DNA to be visible?

I looked at he pGLO through a uv light. Then I put my loop into the pGLO plasmid DNA tube and made sure that the ring was covered with he solution, then mixed it into the +pGLO. I then place the tubes back on ice for 10 minutes and I pushed them under the ice. While my tubes were sitting in the ice I begin Ito label my LB nutrient Agar plates on the bottom. When the 10 minutes is up for the tubes in the ice, I removed them from the ice and put them into a film holder and placed them into a heated water bath for 50 seconds.The settings for the water should be 42 degrees Celsius. When the 50 seconds are up, I placed both tubes back on the ice and had them sit in there for 2 minutes. After the 2 minutes I I removed

We were using a restrictive enzyme to cut the DNA into smaller fragments. For the restriction digest we pipet 4 micrometers of enzyme mix into the bottom of each of our colored tubes making sure to use a new tip for each sample. Next, we capped the tubes and mixed the contents by flicking the tube a little bit with our fingers. After we mixed the contents, we tapped the tube to make sure all the liquid would go to the bottom of the tube. Then, we put the colored tubes in the heating blocks and we let them incubate for 35 minutes at 37 degrees Celsius.

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

Four microcentrifuge tubes were placed in a rack, labeled and numbered, in order to identify the group and the DNA/restriction enzyme that it held. Each of the tubes initially received 10 microliters of reaction buffer. There were two samples of suspect DNA provided along with two restriction enzymes (EcoRI and HindIII). Tubes labeled 1 and 2 received 15 μL of DNA from suspect one while tubes 3 and 4 received 15 μL of DNA from suspect two. Following that, 15 μL of Enzyme 1 (EcoRI) were added to tubes 1 and 3, and 15 μL of Enzyme 2 (HindIII) were added to tubes 2 and 4. (Table 1). The tubes were then gently tapped on the counter to mix the DNA and enzyme solution followed by incubation at 37°C for 45 minutes. After incubation, 5 μL of 10x gel loading dye were added to each of the four tubes of suspect DNA. The tubes were then placed on ice while the gel was under preparation.

After poring the gel, wash the bottle with plenty of running water without draining any excess agarose down the sink. When the gel completely solidifies, transfer it into the electrophoresis chamber. Be sure to place the tray so that the sample wells are near the negative pole. Pour about 275ml of electrophoresis running buffer into the buffer chamber, and adjust the buffer level so the gel is completely submerged below the buffer level. Now, remove the gel comb slowly with a rocking motion until the comb is free of the gel.

Using the electric current, scientists pass the DNA through gel, and as smaller molecules get through gel quicker than those of bigger sizes, DNA molecules get separated according the sizes of the molecules. We utilize the property that large molecules move slower, and DNA is slightly negatively charged(due to phosphate groups), so it will move to the positive pole of the gel.

Gel electrophoresis is a procedure used in laboratories to separate DNA, as well as RNA and proteins. A gel slab is placed in a buffer-filled box and an electrical field is applied. The negatively charged DNA will migrate towards the positively charged side, where it can then be recorded and further analyzed.

Agarose is made from agar or polyacrylamide. With gel electrophoresis you can tell how many different things are put together to make one certain item because this process separates all the other ingredients in the mixture. Also, the macromolecules become coated in a negative charge during gel electrophoresis. The other side of the gel chamber is positive and since opposites attract, the macromolecules are pulled to the other side of the gel separating them. An electrical current is applied to the gel which causes the food coloring dye to migrate and separate through the holes In the gel.

The columns were centrifuged at 15,000 x g for 1 minutes to remove the supernatant. 0.5 mL of Buffer QG was added to each column to remove trace amount of agarose, and 0.75 mL of Buffer PE was added to wash the columns for 5 minutes. Finally, the DNA was eluted with 35 µL of Buffer EB into a clean microcentrifuge tube. The elution buffer was allowed to soak in the membrane for 3 minutes before being centrifuged. Eluted DNA samples were stored at -20

Experiment 4 & 5: PCR did not work for my obtained DNA as there was no evidence of movement of the DNA. PCR did work for lab numbers 33 and 12. The size of my fragment could not be determined and cannot compare to the size that was expected. The negative control was clean as there was no traces of movements.

Electrophoresis can be used to get DNA fingerprint for forensic purposes or to test genes for a particular disease(Gel electrophoresis).The purpose of gel electrophoresis is to picture, identify and distinguish between the molecules that have been processed by other methods such as PCR or enzymatic digestion. Different methods are run through the gel electrophoresis to differentiate between molecules (Gel Electrophoresis - Definition, Purpose and

Nucleic acid molecules which are to be analyzed are set upon a viscous medium, thegel, where an electric field induces the nucleic acids to migrate toward the anode, due to the net negative charge of the sugar-phosphate backbone of the nucleic acid chain. The separation of these fragments is accomplished by exploiting the mobilities with which different sized molecules are able to pass through the gel. Longer molecules migrate more slowly because they experience more resistance within the gel. The DNA fragments of different lengths are visualized using a fluorescent dye specific for DNA, such asethidium bromide. The gel shows bands corresponding to different nucleic acid molecules populations with different molecular weight. Fragment size

Let the test tube sit undisturbed for 2 - 5 minutes. You should begin to see air bubbles form at the boundary line between the ethanol and the filtered fruit solution. Bubbles will form near the top, and you will eventually see the DNA float to the top of the ethanol. Gently insert the stir stick into the test tube. Slowly raise and lower the tip several times to spool and collect the DNA. If there is an insufficient amount of DNA available, it may not float to the top of the solution in a form that can be easily spooled or removed from the tube. However, the DNA will still be visible as white/clear clusters by gently stirring the solution and pushing the clusters around the top. Post-Lab Questions What is the texture and consistency of the DNA DNA is viscous and greasy. Why did we use a salt in the extraction solution High salt makes DNA less soluble in water. In order to dissolve, the water needs to interact with the DNA. Since DNA is quite large, it needs to interact with lots of water for this purpose. When you add salt, the water preferentially interacts with the salt (its small, and can move around in solution easier than the DNA can). This makes it so there is less water available to interact with the DNA and it becomes less soluble. Is the DNA soluble in the aqueous solution or alcohol DNA is less soluble in an alcohol such as isopropanol than it is in water. This is because alcohols are non-polar, whereas water is polar. The polar