It is important to have a high-quality DNA that is free of contaminants such as debris, protein and RNA prior to performing the PCR as well as cloning http://www.mpsciences.com/2021/04/15/gene-synthesis-and-transcription-processes/ or DNA sequencing. 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 for better results.
The initial step in the DNA purification process is to prepare a solution which includes a mix of water and an alkaline buffer. This buffer makes DNA soluble, which means it is able to be separated from the other components of the sample. After the DNA is placed in an alkaline and water solution, it's then treated by chaotropic salts or detergents to destroy cell membranes and nuclei to release DNA (cell lysis). RNase is also added to remove any contaminants in the RNA sample.
The DNA is separated by organic solvents such as chloroform or phenol from the other cellular components, such as proteins and fats. After the DNA is removed from proteins and lipids, it can be extracted using ethanol, or isopropyl alcohol (rubbing alcohol).
Gel electrophoresis and spectrophotometers can be used to determine the purity of DNA. A good quality sample of DNA should have an absorbance ratio of between 260 nm and 280 nm.