June 9, 2024

The Basics of DNA Purification


DNA purification is a crucial element in a number of molecular tests which include PCR as well as qPCR and DNA sequencing. It removes proteins that are contaminating, salts, and other impurities which hinder the downstream process. It also ensures that the desired DNA is present and pure to be used for further analysis. The quality of DNA is determined through spectrophotometry (the ratio of A260 to A280) and gel electrophoresis and a variety of other methods.

In the first step of a DNA purification procedure, the cellular structure will be disrupted using detergents or reagents like SDS in order to release DNA. To further purify DNA, reagents that denature proteins, such as sodium dodecyl sulfate and Ethylene Diamine Tetraacetic Acid (EDTA) can be added to denature them. The proteins are then removed from the nucleic acid solution by centrifugation and then washing. If RNA is found in the sample then it can be further denatured by adding ribonuclease. The nucleic acid is diluted with ice-cold ethanol to isolate it from other contaminants.

Ethanol is a common solvent that can be used to remove salts and other contaminants from nucleic acid samples. Utilizing a standard ethanol concentration allows researchers to compare results across experiments, making it a great choice for workflows that require http://www.mpsciences.com/2021/04/15/gene-synthesis-and-transcription-processes/ high-throughput. Other solvents, like chloroform or phenol can be utilized, however they are more hazardous and require additional steps to avoid cross-contamination. Newer methods can make it easier to complete the DNA purification process using low-ionic-strength ethanol, which has been shown to be equally effective as conventional organic solvents in purifying DNA [26]. This is especially true when combined with a spin column-based extract kit.

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