The Basics of DNA Purification

DNA purification is a vital component of many molecular assays that include PCR, qPCR, and DNA sequencing. It eliminates contaminants like salts, proteins and other impurities that can interfere with downstream processes. It also ensures that the desired DNA is pure and present so that it can be further analysed. The quality of DNA can be evaluated using spectrophotometry, gel electrophoresis and other methods.

In the initial step of a DNA purification process the cell structure will be disrupted with detergents or reagents, such as SDS in order to release DNA. To further purify the DNA, protein-denatured reagents like sodium dodecylsulfate as well as Ethylene diamine tetraacetic acids (EDTA) are added to break down the proteins. They then are removed from the nucleic acid solution using centrifugation and wash steps. If RNA is detected in the sample it can be further denatured by adding ribonuclease. Then, the nucleic acid is diluted with ice-cold ethanol to remove it from other Artificial gene synthesis contaminants.

Ethanol is an everyday solvent that can be used to eliminate salts and other contaminants from nucleic acids samples. Researchers can compare results between experiments using the standard ethanol concentration, which is a great choice for high-throughput workflows. Other solvents, like chloroform or phenol, could be used, but they are more corrosive and require additional steps to avoid cross-contamination. The purification of DNA can be simplified by using ethanol with low ionic strength. This has been shown to be as effective as conventional organic solvents at cleaning DNA. This is especially applicable when used in conjunction with spin column-based extract kits.

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