It is crucial to have high quality DNA that is free of contaminants like proteins, debris, and RNA before performing a PCR, cloning, or DNA sequencing. Purifying DNA is also referred to as DNA isolation and is an essential step in molecular biology. This article will explain the fundamentals of DNA extraction and how to optimize it for better more information results.
The first step in the process of purifying DNA is to prepare a solution which contains a mixture of water and an alkaline buffer. This buffer makes DNA soluble and it is easy to separate from other components in the sample. After the DNA has been placed in an alkaline and water solution, it is then treated with detergents and salts that break down cell membranes and nuclei. This allows the DNA to be released. RNase can be added to the sample in order to remove any contaminating DNA.
DNA is then separated from other cellular components like proteins and lipids by using organic solvents such as phenol and chloroform. After the DNA has been removed from the proteins and lipids, it can be precipitated using ethanol or isopropyl alcohol (rubbing alcohol).
Gel electrophoresis and spectrophotometers can be used to determine the purity of DNA. A high-quality DNA sample must have a ratio of absorbance of 260 nm up to at least 280 nm.