Dna Digestion and Electrophoresis

728 WordsMar 7, 20133 Pages
DNA DIGESTION AND ELECTROPHORESIS In this experiment we will be doing a process called as DNA digestion or also known as restriction digest. A restriction digest is a procedure used in molecular biology to prepare DNA for analysis or other processing. It is sometimes termed DNA fragmentation, scientists Hartl and Jones describe it this way: This enzymatic technique can be used for cleaving DNA molecules at specific sites, ensuring that all DNA fragments that contain a particular sequence have the same size; furthermore, each fragment that contains the desired sequence has the sequence located at exactly the same position within the fragment. The cleavage method makes use of an important class of DNA-cleaving enzymes isolated primarily…show more content…
One of the most important reaction conditions which varies between different restriction enzymes is the salt concentration. Enzyme buffers are specifically formulated to provide the salt concentration for optimal enzyme activity. It is important, therefore, that the correct buffer solution is used for a particular restriction enzyme. [3] For this experiment we also made use of agarose gel electrophoresis, which takes a lot of time. Electrophoresis may be the main technique for molecular separation in today's cell biology laboratory. In spite of the many physical arrangments for the apparatus, and regardless of the medium through which molecules are allowed to migrate, all electrophoretic separations depend upon the charge distribution of the molecules being separated. Electrophoresis can be one dimensional or two dimensional. One dimensional electrophoresis is used for most routine protein and nucleic acid separations. Two dimensional separation of proteins is used for finger printing , and when properly constructed can be extremely accurate in resolving all of the proteins present within a cell. The support medium for electrophoresis can be formed into a gel within a tube or it can be layered into flat sheets. The tubes are used for easy one dimensional separations, while the sheets have a larger surface area and are better for two- dimensional separations. In electrophoresis, proteins are separated on the basis of
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