The Basics of DNA Purification
Before performing a PCR reaction as well as cloning experiments or DNA sequencing, it is crucial to have a high-quality DNA source that is free of contaminants, such as debris, protein and RNA. The process of purifying DNA is known as DNA isolation and is one of the most critical steps in molecular biology. This article will help you understand the basics of DNA extraction and how to optimize it to achieve better results.
The first step of the DNA purification process is to prepare a solution containing an emulsion of water and alkaline buffer. This buffer makes DNA soluble, and it is able to be separated from the other components of the sample. Once the DNA is in an alkaline and water solution, it is then click for source treated by chaotropic salts or detergents to remove cell membranes and nuclei, and release the DNA (cell lysis). RNase may also be added to remove any contaminating RNA from the sample.
The DNA is separated by organic solvents, such as chloroform or phenol from the other components of the cell including fats and proteins. After the DNA is separated from the proteins or lipids it can be precipitated by ethanol or rubbing alcohol.
The quality of the DNA can then be assessed by spectrophotometry or gel electrophoresis. A good quality sample of DNA should have an absorbance range of 250 nm and 280nm.
