Electrophoresis

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Electrophoresis is a separation technique frequently used in the analysis of proteins and nucleic acids. The process known as electrophoresis, involves the migration of particles or molecules (in particular proteins, DNA, and RNA) through an electric field that separates them exclusively on the basis of their size or molecular weight. The direction the molecule moves depends on its charge while the rate of migration is affected by the size, shape, density of the gel and the strength of the applied current (5c). Electrophoresis is a very simple process and relatively quick with a high resolution. In addition electrophoresis is an extremely useful method to estimate the purity of a sample. The technique is also very sensitive to slight variations in molecular weight, size, and even shape of nucleic acids and proteins [1]. Electrophoresis can also be useful when it doesn’t affect the molecule’s structure or denature the protein Cite error: Invalid <ref> tag; invalid names, e.g. too many

Generation of Heat in Electrophoresis Instrumentation

As shown in the diagram, the nucleic acids or proteins are loaded into the wells or depressions at one end on the eletrophoretic medium (also known as a gel). The apparatus also has two electrodes on either side of the eletrophoretic medium. The anode is positively charged while the cathode is negatively charged. When a power source connects the two electrodes the charged particles begin to migrate towards the oppositely charged electrode due to the electric potential field within the media (1).

Electrophoretic Mediums

Detection Techniques

Gel Electrophoresis of DNA and RNA

Gel Electrophoresis of Proteins

Native Electrophoresis

Denaturing Gel electrophoresis

Disadvantage/Advantage

Isoelectric focusing

Two-Dimensional Electrophoresis for Proteins

Two-Dimensional Electrophoresis for DNA and RNA

Blotting Techniques

Advantages & Disadvantages of Electrophoresis

References

  1. Harrison, R. G., Todd, P., Rudge S. R., Petrides, D. P. (2003). Bioseparations Science and Engineering. New York, NY: Oxford University Press.