DNA purification is a crucial element in a variety of molecular tests, such as PCR and qPCR. It removes contaminants, such as proteins, salts and other impurities that can interfere with downstream processes. It also ensures that the desired DNA is clean and available so that it can be used in future analyses. The quality of DNA can be determined through spectrophotometry and gel electrophoresis and various other methods.
The first step in the process of DNA purification is cell lysis. In this process, the cellular structure is broken by reagents or detergents such as SDS to release DNA. To further purify the DNA, reagents with protein denature such as sodium dodecyl sulfate and Ethylene diamine tetraacetic acid (EDTA) are used click for source to denature proteins, and they are removed from the nucleic acid solution using centrifugation and wash steps. If RNA is present in the sample, a ribonuclease treatment can be added to further denature the RNA. The nucleic acids then are concentrated in ice-cold alcohol to isolate them from other contaminants.
Ethanol is a solvent to remove salts and other contaminants from nucleic acids. Using a standardized ethanol concentration allows researchers to compare results across experiments, making it an ideal choice for high-throughput workflows. Other solvents, such as chloroform or phenol, can be used but they are more corrosive and require additional steps to avoid cross-contamination. Modern techniques can speed up the DNA purification process using low-ionic-strength ethanol, which has been shown to be as effective as traditional organic solvents in purifying DNA [2626. This is especially applicable when used in conjunction with spin column-based extract kits.
