Mini Review - Journal of Clinical and Bioanalytical Chemistry (2023) Volume 7, Issue 4
Understanding electrophoresis: Principles and applications.
Department of Chemistry, University of Toronto, Toronto, Canada
- Corresponding Author:
- Robert Johnson
Department of Chemistry
University of Toronto, Canada
Received: 21-July-2023, Manuscript No. AACBC-22-108305; Editor assigned: 22-July-2023, PreQC No. AACBC-22-108305(PQ); Reviewed: 05-Aug-2023, QC No. AACBC-22-108305; Revised: 10-Aug-2023, Manuscript No. AACBC-22-108305(R); Published: 17-Aug-2023, DOI:10.35841/aacbc-7.4.158
Citation: Johnson R. Understanding electrophoresis: Principles and applications. J Clin Bioanal Chem. 2023;7(4):158
Electrophoresis is a fundamental technique in the field of biochemistry, molecular biology, forensics, genetics, and biomedical research, allowing scientists to separate macromolecules such as DNA, RNA, and proteins based on their size, shape, and charge. It's a tool that has played a crucial role in some of the most significant scientific breakthroughs of the 21st century. Electrophoresis, at its most basic, operates on the principle that charged particles will migrate in an electric field. When these particles are placed in a conductive medium (typically a liquid or a gel), and an electric current is applied, they will move towards the electrode that carries a charge opposite to their own. For instance, negatively charged molecules such as DNA will migrate towards the positively charged anode .
The rate at which these particles move is determined by several factors including their charge, size, shape, the strength of the electric field, and the properties of the medium in which they are moving. Generally, smaller particles move faster than larger ones, and more strongly charged particles move faster than less strongly charged ones. Gel Electrophoresis technique is the conductive medium is a gel, most commonly agarose or polyacrylamide. The gel acts as a sort of molecular sieve, allowing smaller molecules to pass through more easily than larger ones. DNA, RNA, and proteins are the typical samples used in gel electrophoresis, with separate processes (like SDSPAGE for proteins) tailored to their unique properties .
Capillary Electrophoresis is the medium is a small capillary tube filled with a conductive liquid. Capillary electrophoresis has the advantage of being faster and more sensitive than gel electrophoresis, and it can provide more detailed information about the molecules being analyzed. Two-dimensional (2D) Gel Electrophoresis is this method combines two different types of electrophoresis. In the first dimension, proteins are separated by isoelectric focusing based on their isoelectric point, i.e., the pH at which the molecules carry no net electrical charge. In the second dimension, the proteins are further separated according to their molecular weight using SDS-PAGE. This results in a high-resolution separation of complex protein mixtures .
Pulsed-field gel electrophoresis is specifically designed for the separation of large DNA molecules by applying electrical pulses that change direction. It’s widely used in molecular biology for the typing of bacteria for epidemiological studies.
Immunoelectrophoresis is this method combines electrophoresis and immunoassay to characterize and quantify proteins, particularly antibodies and antigens. It has applications in both clinical diagnostics and laboratory research.
Electrophoresis is used to analyze DNA and RNA fragments, helping to identify gene mutations related to genetic disorders. It's a cornerstone of the Human Genome Project, which mapped every gene in the human genome. In biochemistry and molecular biology, electrophoresis is employed to analyze proteins' structure and function. It can reveal the presence or absence of specific proteins, helping to diagnose various diseases. In forensic science, DNA electrophoresis plays a vital role. It is used to compare DNA samples from crime scenes with those of suspects or to establish biological relationships in paternity testing. Electrophoresis is used to test the purity of drugs and to separate and identify different compounds within a mixture .
Electrophoresis continues to evolve as scientists develop new variations and improvements. For instance, two-dimensional electrophoresis allows for a more complex analysis of proteins, separating them by both size and charge. Microchip electrophoresis, another recent advancement, miniaturizes the process onto a tiny chip, increasing speed and efficiency, electrophoresis is a powerful tool in scientific research, offering a versatile method for analyzing macromolecules. As technologies continue to advance, the future of electrophoresis promises to unlock even more exciting opportunities for discovery and understanding .
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