DNA purification is a crucial element in a number of molecular tests which include PCR, qPCR, and DNA sequencing. It eliminates contaminants such as salts, proteins, and other impurities which can interfere with downstream processes. It also ensures that the desired DNA is pure and present in order to be used for further analysis. The quality of DNA is determined by spectrophotometry (the ratio of A260 to A280) as well as gel electrophoresis and many other methods.
The initial step in the process of DNA purification is cell lysis. In this process, the cell structure is destroyed by reagents or detergents such as SDS to release DNA. To further purify DNA, protein-denatured reagents like sodium dodecyl sulfate and Ethylene diamine tetraacetic acids (EDTA) are used to denature proteins, and they are removed from the nucleic acid solution using centrifugation and washing steps. If RNA is detected in the sample it can be further denatured by adding ribonuclease. The nucleic acids are then concentrated in ice-cold water to separate them from other contaminants.
Ethanol is solvent to eliminate salts and other contaminants from nucleic acids. Using a standardized concentration of ethanol lets researchers compare the results of different studies, making it a good option for workflows that require high-throughput. Other solvents, such as chloroform or phenol, can be used, however, they are more toxic and require additional steps to prevent cross-contamination. Newer techniques can simplify the DNA purification process using ethanol with low-ionic strength, which has been proven to be as efficient as the conventional organic https://mpsciences.com/2021/04/01/types-of-science-products-available/ solvents when purifying DNA [26]. This is especially the case when paired with spin column-based extract kits.