Opinion Article - Journal of RNA and Genomics (2023) Volume 19, Issue 1
Electrophoresis for investigating the diversity of biomolecules.
Department of Behavioral Sciences, Leiden University, Leiden, The Netherlands
- Corresponding Author:
- Marni Ollerton
Department of Behavioral Sciences,
Leiden, The Netherlands
Received: 02-Jan-2023, Manuscript No. RNAI-23-95167; Editor assigned: 04-Jan-2023, Pre QC No. RNAI-23-95167(PQ); Reviewed: 19-Jan-2023, QC No.RNAI-23-95167; Revised: 27-Jan-2023, Manuscript No.RNAI-23-95167(R); Published: 03-Feb-2023, DOI:10.4172/2591-7781.1000136.
Citation: Ollerton M. Electrophoresis for investigating the diversity of biomolecules. J RNA Genomics 2023;19(1):1-10.
Electrophoresis is a powerful technique widely used in molecular biology, biochemistry, and analytical chemistry to separate and analyze charged particles, such as proteins, nucleic acids, and other biomolecules, based on their size, shape, charge, and mobility in an electric field. It has become an essential tool in modern scientific research and has found applications in various fields, including genomics, proteomics, forensic science, clinical diagnostics, and drug discovery.
Electrophoresis relies on the movement of charged particles in an electric field, with the rate of migration influenced by the charge, size, shape, and mobility of the particles. The key components of an electrophoresis setup, including the gel matrix, buffer system, and power supply, will be discussed in detail. Different types of electrophoresis techniques, such as agarose gel electrophoresis, Polyacrylamide Gel Electrophoresis (PAGE), capillary electrophoresis, and isoelectric focusing, will be introduced, along with their advantages, limitations, and applications.
Types and Applications of Electrophoresis
The various types of electrophoresis and their applications in different fields of science. Agarose gel electrophoresis is commonly used for the separation of DNA and RNA fragments, while PAGE is widely employed for protein analysis. Capillary electrophoresis is a high-resolution technique used for the separation of small molecules, including nucleotides, amino acids, and carbohydrates. Isoelectric focusing is a specialized technique used for the separation of proteins based on their isoelectric point (pI). Additionally, other advanced forms of electrophoresis, such as two-dimensional electrophoresis, native gel electrophoresis, and immunoelectrophoresis, will be discussed, along with their specific applications in research and diagnostics.
Electrophoresis techniques and procedures
The techniques and procedures involved in performing electrophoresis experiments. It will cover the preparation of the gel matrix, including the formulation of agarose and polyacrylamide gels, and the casting of gel plates. The preparation of the samples, including DNA, RNA, proteins, and other biomolecules, will be discussed, along with the loading and running of the samples on the gel. The composition and preparation of the electrophoresis buffer, including the pH, ionic strength, and additives, will also be covered in detail. The optimization of electrophoresis conditions, such as voltage, temperature, and run time, will be discussed, along with tips for common issues encountered during electrophoresis experiments.
Electrophoresis is a widely used technique in biochemistry, molecular biology, and analytical chemistry for the separation and analysis of charged molecules based on their migration in an electric field. Over the years, electrophoresis has evolved and continues to advance, leading to emerging trends and future directions in the field. Some of the key emerging trends and future directions in electrophoresis include:
Microscale and nanoscale electrophoresis: Miniaturization of electrophoresis systems has gained significant attention in recent years. Microscale and nanoscale electrophoresis techniques offer advantages such as faster separations, reduced sample and reagent consumption, and enhanced sensitivity. Microfluidic and lab-on-a-chip electrophoresis systems are being developed for applications in point-of-care diagnostics, DNA sequencing, proteomics, and drug discovery.
High-throughput electrophoresis: Automation and highthroughput screening are driving advancements in electrophoresis. High-throughput electrophoresis systems are being developed to analyze large numbers of samples simultaneously, allowing for rapid screening and analysis of biological samples in various applications, including drug discovery, genomics, and proteomics.
Capillary electrophoresis (CE): Capillary electrophoresis, which uses narrow-bore capillaries filled with an electrolyte solution, is gaining popularity due to its high separation efficiency and ability to analyze a wide range of analytes. Recent advancements in CE include the development of new capillary coatings, improved sample injection techniques, and the integration of CE with other analytical techniques, such as mass spectrometry and fluorescence detection.
Electrophoresis in novel media: Traditional electrophoresis techniques use gels or capillaries filled with electrolyte solutions as the separation media. However, there is growing interest in using novel media, such as nanomaterials, ionic liquids, and polymer matrices, to enhance the separation efficiency and selectivity of electrophoretic separations. These alternative media offer unique properties that can be exploited for improved separation of complex samples.
Online detection and automation: Integration of electrophoresis with online detection techniques, such as mass spectrometry, fluorescence, and electrochemical detection, is becoming increasingly important for real-time monitoring and analysis of analytes during electrophoresis separations.
Automation of electrophoresis systems through robotics, microfluidics, and artificial intelligence is also gaining momentum, allowing for improved reproducibility, precision, and efficiency of electrophoretic separations.
Novel applications of electrophoresis: Electrophoresis is finding new applications beyond traditional areas, such as genomics and proteomics. For example, electrophoresis is being used for the separation and analysis of extracellular vesicles, circulating tumor cells, and other rare cell populations for diagnostics and therapeutics. Electrophoresis is also being explored for the separation and analysis of non-traditional analytes, such as nanoparticles, exosomes, and liposomes, in fields such as drug delivery, environmental science, and nanotechnology.
Advances in data analysis and bioinformatics: As electrophoresis techniques generate large amounts of data, advances in data analysis and bioinformatics are becoming essential for extracting meaningful information from electrophoretic data. Machine learning, data mining, and statistical methods are being employed for data analysis, pattern recognition, and automation of data interpretation in electrophoresis.