Using Gel Electrophoresis And Dna Fingerprinting To Analyze

For this experiment, we used two restriction enzymes called Ava II and Pvu II. Four tubes were needed for the experiment as well. Each microtubule was labeled accordingly, A, B, and C. Using Micropipettes, we added 2 microliters of pUC19 DNA into each tube. To make sure the DNA was on the bottom of the tube, we tapped each tube on the lab bench. Each tube had its own specific amount of different solutions added on, however the final volume for each tube was 30 microliters. Tube A had, 24 microliters of DI water, 3 microliters of the buffer, and 1 microliter of the Ava II enzyme. Tube B had, 24 microliters of DI water, 3 microliters of the buffer, and 1 microliter of the Pvu II enzyme. Tube C had, 23 microliters of DI Water, 3 microliters of the buffer, 1 microliter of Ava II, and 1 microliter of Pvu II. The last tube C had, 25 microliters of DI water, 3 microliter of the buffer, and no enzymes. Each solution was added according to how it is written here. After we finished adding the solutions to each tube, we tapped the tubes on the lab bench to make sure the

Figure 1 Gel Electrophoresis for Replication Taster PTC. The gel is composed of an ethidium bromide stained 3% agarose gel demonstrating DNA fragments which were a depiction of PCR amplification. The agarose gel contains nine loading samples, including from left to right, the MW marker lane 1 precision mol mass standard, lane 2 TB undigested PTC (5µl of DNA, 5µl of master mix P, and 2.5µl of loading dye), lane 3 TB digested PTC (5µl of DNA, 5µl of master mix P, 2µl Fnu4HI, and 3µl of loading dye), lane 4 TB A(L)DH G (10µl DNA, 10µl master mix G, and 5µl loading dye), lane 5 TB A(L)DH A (10µl DNA, 10µl master mix A, and 5µl loading dye), lane 6 MG undigested PTC (5µl of DNA, 5µl of master mix P, and 2.5µl of loading dye), lane 7 MG digested PTC (5µl of DNA, 5µl of master mix P, 2µl Fnu4HI, and 3µl of loading dye), lane 8 MG A(L)DH G (10µl DNA, 10µl master mix G, and 5µl loading dye), lane 9 MG A(L)DH A (10µl DNA, 10µl master mix A, and 5µl loading dye).

DNA samples can be extracted from hair, blood and skin. Siblings have different DNA fingerprints because everyone has 23 pairs of chromosomes and for each pair one is one of your mother’s chromosomes and the other is your father’s chromosomes. Other than identical twins no other people have exactly the same DNA. DNA fingerprinting is used every day to determine whose parents or siblings are whose, it is used in crime scenes to determine who was at the scene, can be used to determine where a certain inherited gene is inherited from and it can be used to identify a body that is deceased. During electrophoresis an electric current passes through the agarose gel, therefore moving the DNA samples through the gel. The smaller the DNA fragment the faster it moves through the gel. The finished product will look like a series of bands, some will match up and some won’t. The different bands in electrophoresis represent different gene fragments.

This however, was unsuccessful, as the restriction digestion created fragments of many different lengths, resulting in smears at different lengths on the gel electrophoresis due to the restriction enzyme recognition sites being present in-between the telomeric repeat sequences, the analysis was thus

The first agarose gel provided insight into whether or not Zassy’s genomic DNA had been cut by the Sal I HF (NEB) restriction enzyme. As seen in Figure 1, Zassy’s cut DNA, in lane 3 left clear fragments between the 3 kb and 0.5 kb areas. Zassy’s uncut DNA, in lane 2, left a clear and bright band above the 10 kb marker. Therefore, the gel confirmed that Zassy’s DNA was successfully cut with Sal I because there was a significant difference the cut and uncut DNA that was run through the gel. There was an additional lane of cut DNA because another student adopted Zassy in the early stages of this experiment. Nevertheless, the clear distinction between lanes 2 and 3 allowed the experiment to continue on to purification of the DNA.

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.

The gel was covered with a buffer and then six samples labeled A-F were deposited into the wells using a micropipette. Three of these samples (A, B, C) were control samples to compare to D, E, and F (the mother, child, and father). The safety cover was placed on the unit and then brought to a power source. The leads were connected the chamber and left for approximately 20 minutes. The agarose gel was removed from the tray and placed onto a sheet of plastic wrap. An Ethidium Bromide card was placed face down onto the gel to stain for approximately 5 minutes. Finally, the card was removed and the gel was placed on top of a UV light. The samples were pushed towards the center due to opposite electric charges. Agarose gel separates the DNA samples by the way they were cut. The restriction enzyme MST II cuts the DNA strand at CC/TNAGG where N is any nucleotide base. If the enzyme recognizes this, it is cut. If it does not recognize it then the strand is left whole. We were able to observe the DNA strands due to them being dyed and placed over a UV light. The control samples were utilized so that the other samples could be compared to test for their genotype. The data was analyzed in this way to differentiate between the different genotypes and the number of bars they

A restriction enzyme is an enzyme that acts as a catalyst that can cut a strand of DNA in a particular place.

An important part of this is which enzymes will work in the same buffer. Using multiple restriction enzymes is necessary because without multiple enzymes it would impossible to determine the specific sequence of the DNA strand. Once you have determine which restriction enzymes you want to use, you will then set up the digest. You will need several 1.5 mL tubes (at least 7 tubes if you are using 3 enzymes). In each tube you need to add some of your DNA sample. You will then add different combination of restriction enzymes to each tube. The typical order of combination of enzymes would be 3 tubes that have one enzyme each, 3 tubes that have 2 enzymes each and one tube that has all the enzymes. It is necessary to have different combinations of the restriction enzymes so that you get a better idea of how the enzymes cut in relation to each other. Then you will add buffer, BSA, and water. Once the tubes are mixed they have to be incubated so that the restriction enzymes have time to cut the DNA into fragments. After the incubation the tubes of DNA fragments are ran through a gel electrophoresis, which allows you to visualize the results of the

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.

The purpose of the DNA restriction and electrophoresis lab was to first become familiar with the properties of restriction enzymes and discover that they, along with agarose gel electrophoresis, are used to characterize DNA molecules. Restriction enzymes are used to make cuts or join together DNA fragments. The cuts result in either staggered cleavage, or blunt cleavage. Staggered cleavage results when the breaks are offset. This cleavage usually results in sticky end being produced because for each DNA end that is produced there are extensions with unpaired nucleotide bases. Blunt cleavage results when the breaks in the phosphodiester bonds are right across from each other.

Restriction enzymes cut DNA at certain sites to create multiple DNA fragments. Restriction enzyme HindIII has known DNA fragment lengths and recognition sites when digesting lambda DNA, while the lambda DNA recognition site for restriction enzyme XhoI is unknown. The goal of this study is to determine the lambda recognition site of XhoI by comparing a HindIII digest and a HindIII and XhoI double digest on an electrophoresis gel. The HindIII digest had a band at 9.4 kb, but this band was not visible in the double digest, therefore we concluded the recognition site for XhoI was around 9.4kb. There were also two additional DNA

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 is the method of separating and observing macromolecules (DNA, RNA, and proteins) and their fragments. The conditions that allow macromolecules to separate during gel electrophoresis include their charges and sizes. Restriction enzymes split DNA molecules in certain places. In gel electrophoresis, an electric current moves DNA molecules across a slab of agarose gel or a polysaccharide medium, and the polysaccharide medium or the medium with sugar molecules then withholds the DNA molecules as they are conveyed by electric currents. The phosphate groups in DNA molecules’ backbone possess a high negative charge. When DNA molecules are put into a field of electric charge, the high negative charge of DNA molecules force the molecules to run to the positive or opposite end of the agarose gel. Because agarose gel contains small molecules of sugar molecules, large molecules of DNA struggle migrating towards the positive end of the agarose gel because of their bulkiness. Whereas small molecules of DNA have little problem moving or move faster towards the positive end of the agarose mesh because they do not bump into sugar molecules as often as large DNA molecules do. What is more, the main setup of gel electrophoresis is that the gel will be submerged a in buffer

Deoxy ribonucleic acid (DNA) is a nucleic acid that contains genetic instructions used to grow and develop all the organisms. The main role of the DNA molecule is the long-term storage of genetic information. DNA is often compared to a set of maps, because it contains the instructions needed to make all other components of the cell, such as protein molecules and RNA (Ribonucleic Acid). DNA fragments carrying genetic information are called genes, but other segments in the DNA chain also have construction goals or are involved in regulating the use of this genetic information.