The Basics of DNA Purification

DNA purification is a vital element in a variety of molecular tests, including PCR and qPCR. It removes contaminating proteins as well as salts and other impurities that can hinder the downstream process. It also ensures the desired DNA is clean and present in a way that it can be used for further analyses. The quality of DNA can be determined using spectrophotometry, electrophoresis on gels and other methods.

The first step in a DNA purification procedure is cell lysis, in which the cellular structure is disrupted with detergents or reagents such as SDS to release DNA. To further purify DNA, reagents that alter proteins, such as sodium dodecyl-sulfate or Ethylene Diamine Tetraacetic Acid (EDTA) can be added to denature them. The proteins are then removed from the nucleic acids solution by centrifugation and washing. If there is RNA present in the sample, a ribonuclease treatment can be added to further denature the RNA. The nucleic acids are concentrated in ice-cold alcoholic to separate them from other contaminants.

Ethanol is a solvent to remove salts or other contaminants from nucleic acids. Researchers can evaluate the results of different experiments by using the standard ethanol concentration, which is a good option for high-throughput workflows. Other solvents, including chloroform and phenol, can be used, but they are more hazardous and require additional steps to prevent cross-contamination. The purification of DNA can be made simpler by using low ionic strength ethanol. It has been proven to be as effective as conventional organic solvents for eliminating DNA. This is especially true when combined with spin column extract kits.

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