It is essential to have high-quality DNA that is free of contamination such as protein, debris and RNA prior to performing a PCR or cloning procedure, or DNA sequencing. The process of purifying DNA is known as DNA isolation, and is one of the most vital steps in molecular biology. In this article, you’ll learn the basics of DNA purification and how to optimize your DNA extraction strategies to get more efficient results.
The initial step in the DNA purification process is to prepare a solution which contains a mixture of water and an alkaline buffer. This buffer makes the DNA more soluble so that it will easily separate from the other components of the sample. After the DNA is placed in an alkaline and water solution, it’s treated by chaotropic salts or detergents to destroy cell membranes and nuclei and release DNA (cell lysis). RNase can be added to the sample in order to remove any DNA that has been contaminated.
DNA https://mpsciences.com/2021/04/08/different-types-of-pcr-reagents/ is then separated from other cell components such as proteins and lipids with the help of organic solvents like phenol and chloroform. After the DNA has been removed from lipids or proteins it can be precipitated by alcohol or ruby alcohol.
Spectrophotometry and gel electrophoresis can be used to determine the purity of DNA. A good quality DNA sample should have a ratio of absorbance between 260 and at least 280 nm.