It is essential to have high quality DNA that is free of contamination such as proteins, debris and RNA prior a PCR as well as cloning or DNA sequencing. The process of purifying DNA is referred to as DNA isolation. It is one of the most crucial steps in molecular biology. This article will explain the basics of DNA extraction and how to optimize it to achieve better results.
The initial step of the DNA purification procedure is to make a solution that consists of the blog mixture of alkaline buffer and water. This buffer makes the DNA soluble so that it is able to be separated from other components of the sample. Once the DNA has been placed in an alkaline and water solution it is treated with detergents and salts that break down cells’ membranes and nuclei. This releases the DNA. RNase is also added to remove any contamination of RNA from the sample.
DNA is then separated from other cellular components like proteins and lipids with the help of organic solvents like phenol and chloroform. Once the DNA is removed from the proteins or lipids it can be precipitated with alcohol or ruby alcohol.
Spectrophotometry and electrophoresis may be used to determine the purity of DNA. A good quality DNA sample should have an absorbance range of 220 nm to 280 nm.
