Before making a PCR reaction or cloning experiment, or even DNA sequencing it is critical to have high-quality DNA which is free of contaminants such as debris, proteins and RNA. Purifying DNA is also known as DNA Isolation, and is a vital step in molecular biology. This article will explain the basics of DNA extraction and how to improve it for better results.
The initial step of the DNA purification process is to make a solution consisting of an emulsion of water and alkaline buffer. This buffer makes DNA soluble, so it is easily separated from other components of the sample. Once the DNA is in an alkaline and water solution, it is then treated by chaotropic salts or detergents to dissolve cell membranes as well as nuclei and release DNA (cell lysis). RNase can be added into the sample to remove any DNA that is contaminating.
DNA is then separated from other cell components such as proteins and lipids by using organic solvents such as chloroform and phenol. After the DNA has been removed from lipids or proteins and lipids, it can be precipitated using ethanol or ruby alcohol.
The purity of the DNA can then be verified using spectrophotometry or gel electrophoresis. A high-quality sample of https://mpsciences.com/2021/04/23/dna-purification-processes-for-different-applications/ DNA should have an absorbance value between the 260-nm range and 280-nm range.