Electrophoresis

Electrophoresis is a technique used to separate macromolecules in a fluid or gel based on their charge, binding affinity, and size under an electric field. Anaphoresis is the electrophoresis of negative charge particles or anions whereas cataphoresis is electrophoresis of positive charge ions or cations.

Electrophoresis is a general term that describes the migration and separation of charged particles (ions) under the influence of an electric field. An electrophoretic system consists of two electrodes of opposite charge (anode, cathode), connected by a conducting medium called an electrolyte.

Gel electrophoresis is a technique used to separate DNA fragments according to their size. DNA samples are loaded into wells (indentations) at one end of a gel, and an electric current is applied to pull them through the gel. DNA fragments are negatively charged, so they move towards the positive electrode.

It is a laboratory method used to separate mixtures of DNA, RNA, or proteins according to molecular size. In gel electrophoresis, the molecules to be separated are pushed by an electrical field through a gel that contains small pores.

An electric current is used to move molecules to be separated through a gel. Pores in the gel work like a sieve, allowing smaller molecules to move faster than larger molecules.

In this manner, DNA fragments in a solution are separated on the basis of size. There are several basic steps to performing gel electrophoresis that will be described below; 1) Pouring the gel, 2) Preparing your samples, 3) Loading the gel, 4) Running the gel (exposing it to an electric field) and 5) Staining the gel.

The serum protein electrophoresis (SPEP) test measures specific proteins in the blood to help identify some diseases. Proteins are substances made up of smaller building blocks called amino acids. Proteins carry a positive or a negative electrical charge, and they move in fluid when placed in an electrical field.

Two applications of DNA profiling are: Paternity testing (comparing DNA of offspring against potential fathers) Forensic investigations (identifying suspects or victims based on crime-scene DNA).

The negatively charged DNA molecules migrate towards the positive charge under the influence of constant current, thus the separation depends on the mass and charge of DNA. The DNA molecules are forced to move through the agarose gel pores.

If you use water instead of buffer for the gel or running buffer. Agarose gels are cast and run using buffer. If you do use water, your gel will melt shortly after applying voltage to the electrophoresis unit.

In agarose gel electrophoresis, EDTA is added in buffer for chelating the magnesium ions which are cofactors for DNA nucleases. Hence, activity of DNA nucleases that may be present is inhibited, and DNA is protected from degrading by DNA nucleases.

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Nicola B
Editorial Team
Journal of  Biochemistry and Biotechnology