It is important to have high-quality DNA that is free of contamination such as proteins, debris and RNA prior to carrying out a PCR, cloning, or DNA sequencing. Purifying DNA is also known as DNA isolation and is an essential step in molecular biology. This article will help you understand the basics of DNA extraction and how to improve it for better results.
The initial step of the DNA purification process is to prepare a solution that contains an amalgamation of water and alkaline buffer. This buffer makes the DNA soluble and so that it is able to be separated from other components of the sample. After the DNA has been placed in an alkaline and water solution, it’s cleaned with detergents as well as Chaotropics salts in order to break up cell membranes and nuclei. This releases the DNA. RNase can be added into the sample to remove any DNA contamination.
DNA is separated from other cellular components like proteins and lipids with the help of organic solvents like chloroform and phenol. After the DNA is separated from lipids or proteins, it can be precipitated using ethanol or ruby alcohol.
The purity of the DNA can then be confirmed using spectrophotometry, or gel electrophoresis. A good quality sample of DNA should have an absorbance range of 220 nm to 280 nm.
https://mpsciences.com/2021/04/08/different-types-of-pcr-reagents/