DNA purification is a crucial element in a number of molecular tests such as PCR or qPCR as well as DNA sequencing. It removes contaminants, such as salts, proteins and other impurities that can interfere with downstream processes. It also ensures that the desired DNA is present and pure in order to be further analysed. The quality of DNA can be determined by spectrophotometry, electrophoresis on gels and other methods.
The first step of a DNA purification procedure is cell lysis. This is when the cellular structure is broken by reagents or detergents 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 acid solution by centrifugation and washing. If RNA is present in the sample, a ribonuclease treatment can be added to further denature the RNA. The nucleic acids are then concentrated in ice-cold water to separate them from other contaminants.
Ethanol is an everyday solvent that can be used to remove salts and other contaminants from nucleic acids samples. A standard concentration of ethanol allows researchers to compare the results of different studies, making it a good option for workflows that require high-throughput. Other click this link now solvents, like chloroform or phenol, can be used, however, they are more hazardous and require additional steps to prevent cross-contamination. The purification of DNA can be simplified by using ethanol with low ionic strength. It has been proven to work just as conventional organic solvents for eliminating DNA. This is particularly true when combined with spin column extract kits.
