Research Paper on Gel Electrophoresis Essay
Gel Electrophoresis is a technique in molecular biology and biochemistry, which is used for the separation of proteins and nucleic acids, based on the motion of charged biological macromolecules in a constant electric field. Polyacrylamide gel separation is due to differences in molecular charge and separated differences of molecular weights and the configuration of molecules. There are so-called non-denaturating or native PAGE electrophoresis (where the shared biological macromolecules are in the native state during electrophoresis) and denaturing polyacrylamide gel electrophoresis (where the samples were denatured beforehand in the case of nucleic acids using short heating the sample with glyoxal or formamide, to denature the proteins normally boiling the samples used in buffer containing a strong ionic detergent (usually sodium dodecyl sulphate) and the agent that disrupts the quaternary structure of the protein by breaking disulfide bridges between protein globules inside polypeptide chain – beta-mercaptoethanol).
During the process of denaturing by PAGE electrophoresis, molecules are stored in the denatured state by the presence of chaotropic agents in the gel (typically urea) in the case of the PAGE electrophoresis of nucleic acids and proteins through the presence of the ion (e.g. sodium dodecyl sulfate, cetyltrimethylammonium bromide) and non-ionic (e.g., tween-20) detergents.
In the case of polyacrylamide gel electrophoresis of proteins method the modification of Laemmli is generally used.
Also polyacrylamide gel electrophoresis is used to separate the short nucleic acid fragments, e.g., DNA electrophoresis, in cases such as Sanger sequencing. Additionally, SDS-PAGE electrophoresis is used in methods for visualizing the restriction fragment length polymorphismand polymerase chain reaction method.
There is also so-called disc-electrophoresis (from discontinuous), where there is pH gradient in the gel during the electrophoretic separation of proteins at the interface between the concentrating and separating gels, thereby achieving better separation of protein molecules.
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