DNA purification is a critical step in many molecular tests such as PCR and qPCR. It eliminates harmful proteins salts, proteins, and other impurities which hinder the downstream process. It also ensures the desired DNA is clean and in good condition so that it can be used in further studies. The quality of DNA can be determined through spectrophotometry (the ratio of A260 to A280), gel electrophoresis, and many other methods.
In the beginning of a DNA purification process, the cellular structure will be disrupted by using https://mpsciences.com/2021/04/23/dna-purification-processes-for-different-applications/ detergents or reagents like SDS to release DNA. To further remove DNA, reagents that are able to denature proteins like sodium dodecyl sulfate or Ethylene Diamine Tetraacetic Acid (EDTA) can be added to denature them. The proteins are removed from the nucleic acids solution by centrifugation and then washing. If RNA is present in the sample, a ribonuclease treatment could be added to further denature RNA. The nucleic acid is reacted with ethanol that has been cooled to remove it from other contaminants.
Ethanol can be used as solvents to remove salts or other contaminants from nucleic acids. Using a standardized concentration of ethanol allows researchers to examine the results of various studies, making it a great choice for workflows that require high-throughput. Other solvents, such as chloroform or phenol, can be used, but they are more harmful and require additional steps to prevent cross-contamination. Newer techniques can facilitate the process of purifying DNA by using low-ionic-strength ethanol that has been shown to be as efficient as traditional organic solvents for purifying DNA [2626. This is particularly the case when paired with a spin column extraction kit.
