Before making a PCR reaction, cloning experiment or DNA sequencing it is critical to have high-quality DNA that is free of contaminants such as debris, protein and RNA. 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 learn the basics of DNA purification as well as how to optimize your DNA extraction methods to get better results.
The initial step of the DNA purification procedure is to prepare a solution that contains an emulsion of alkaline buffer and water. This buffer makes the DNA more soluble so that it will easily separate from other components of the sample. Once the DNA has been placed in an alkaline and water solution, it is then treated with detergents and Chaotropics salts in order to break up cell membranes and nuclei. This allows the DNA to be released. RNase may also be added to eliminate any contamination RNA from the sample.
DNA is separated from other cellular components such as proteins and lipids with the help of organic solvents such as chloroform and phenol. After the DNA is removed from the proteins or lipids it can be precipitated by alcohol or rubbing alcohol.
The quality of the DNA sample can be assessed by https://mpsciences.com/2021/04/23/dna-purification-processes-for-different-applications/ spectrophotometry or gel electrophoresis. A good quality DNA sample should have a ratio of absorbance of 260 nm up to the range of 280 nm.
