What are the different methods of nucleic acid extraction used in molecular diagnostics?

In 1953, an important discovery of the double helix structure of deoxyribonucleic acid (DNA) by James Watson, Rosalind Franklin and Francis Crick made huge waves in medical science. The study of DNA structure has since inspired many advancements in genetic science, especially in the field of molecular diagnostics or testing.

Over the last two decades, research on the human genome has expanded into fields like as genetic analysis and engineering, antibody treatments, gene therapies, and genome sequencing, all of which are pillars of modern biotechnology.

The helix structure is made up of two long chains of single nucleic acid molecules, commonly known as nucleotides, that are twisted around one another. There are two types of nucleic acids:

• Deoxyribonucleic acid (DNA) and

• Ribonucleic acid (RNA) acid.

In living creatures, DNA and RNA molecules serve as a repository for crucial genetic information. DNA is made up of complementary double nucleotide strands. One nucleotide strand of RNA is created from a single strand of DNA.

The study of nucleic acid molecule extracts shows the nature of the stored information, how it is transported and reproduced, and how it influences, controls, and drives the development and operation of living organisms' chemical processes. Furthermore, clinical research on nucleic acid molecules has led to the identification of disease-related DNA sequences associated with hereditary illnesses, cancer, and infectious diseases.

More crucially, several molecular testing techniques, such as polymerase chain reactions (PCR), Sanger, and next-generation sequencing, require isolated DNA and RNA input. As a result, nucleic acid extraction methods have a specific place in molecular biology.

Helvetica Health Care (HHC) outlines what nucleic acid extraction is and the numerous methods that are commonly employed in biomedical research in this article.

What is nucleic acid extraction?

Nucleic acid extraction is a process which involves the following steps.

1) Disruption of the cells and tissues – also called Cell Lysis.

2) Isolation or purification of the nucleic acids of proteins, lipids, and other impurities through solution based, solid-phase or magnetic methods.

3) Moving or transferring the nucleic acids into a buffer solution or water for preservation without affecting further research.

What are the different nucleic acid extraction methods used in medical research?

The following sections provide an overview of some of the most often utilized nucleic acid extraction techniques.

Cesium chloride gradient centrifugation (with ethidium bromide)

Density gradient centrifugation, which has been used in clinical research since the 1950s, exploits changes in density and buoyancy between caesium ions and water, as well as Ethidium Bromide intercalation, to impede DNA transcription, repair, replication, and recombination. The procedure enables for the separation of various DNAs and aids in the recovery of DNA with a high yield.

Solid-Phase Extraction

Solid phase extraction, often known as the spin column method, is based on attaching nucleic acids to solid supports. DNA is extracted rapidly and efficiently utilising a spin column powered by centrifugal force, which eliminates the need for previous liquid extraction methods and poor phase separation. Silica is the most commonly utilised solid support for nucleic acid extraction.

Magnetic Bead Method

The magnetic bead method, a variant of the solid-phase extraction technique, is a handy way to isolate a sample's DNA from proteins and other undesirable biological components that bind on negatively charged magnetic beads. Scientists and lab professionals can avoid repeating the centrifugation, vacuum filtration, and column separation stages for washing, elution, and organic solvents by using this method.

Alkaline extraction

The initial stage in plasmid DNA separation and purification is alkaline extraction. The significance of this approach arises from its ability to extract the DNA and proteins that are ordinarily bound to chromosomal DNA. Alkaline extraction is intended for use in molecular cloning processes that require chromosomal DNA extraction in order to modify plasmid DNA.

Phenol–Chloroform Extraction

This solution-based extraction approach is also known as the Guanidinium Thiocyanate-Phenol-Chloroform nucleic acid extraction. The approach entails combining salt guanidinium thiocyanate (an aqueous solution) with phenol and chloroform, which are solvents for effective RNA purification.

Cetyltrimethylammonium Bromide Nucleic Acid Extraction

This method is used to extract DNA from plant and, in certain situations, food samples. It enables excellent removal of plant-based detritus from nucleic acid prior to further purification.

Chelex DNA Extraction

The Chelex technique, which is mostly used in forensics, may extract DNA from a variety of sources, including bloodstain cards, buccal swabs, and hair, by utilising a resin that binds to common PCR (polymerase chain reaction) inhibitors. Despite the slightly raw output, it safely protects DNA for PCR-based forensic investigation applications.

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