The Basics of DNA Purification
It is essential to have high-quality DNA that is free of contaminants like protein, debris and RNA prior to carrying out the PCR or cloning process, or DNA sequencing. The process of purifying DNA is referred to as DNA isolation. It is one of the most vital steps in molecular biology. In this article, you will discover the fundamentals of DNA purification, and how to optimize your DNA extraction strategies for better results.
The initial step of the DNA purification procedure is to prepare a solution containing an amalgamation of water and alkaline buffer. This buffer makes DNA soluble, which means it is easily separated from other components in the sample. After the DNA is placed in an alkaline and water solution, it’s then treated by chaotropic salts or detergents to remove cell membranes and nuclei, and release the DNA (cell lysis). RNase may also be added to eliminate any contamination of RNA from the sample.
DNA is later separated from other cellular components like proteins and lipids with the help of organic solvents such as phenol and chloroform. Once the DNA is removed from lipids or proteins then it can be precipitated using ethanol or rubbing alcohol.
The quality of the DNA can be determined by spectrophotometry or gel electrophoresis. A high-quality DNA sample should have a ratio of absorbance of 260 nm up to the range of 280 nm.