The Basics of DNA Purification

It is crucial to have high-quality DNA that is free of contamination such as protein, debris, and RNA before performing a PCR as well as cloning or DNA sequencing. The process of purifying DNA is referred to as DNA isolation and is considered to be one of the most important steps in molecular biology. In this article you will learn the basics of DNA purification, and how to optimize your DNA extraction techniques for more efficient results.

The first step in the DNA purification process is to prepare a solution that is composed of water and an alkaline buffer. This buffer makes DNA soluble, which means it is easily separated from other components in the sample. Once the DNA is in an alkaline and water solution, it’s treated with chaotropic and detergents to dissolve cell membranes as well as nuclei and release DNA (cell lysis). RNase may also be added to remove any contaminating RNA from the sample.

The DNA is separated by organic solvents such as chloroform and phenol from other cellular components like proteins and fats. After the DNA has been removed from the proteins and lipids, it can be extracted using ethanol or isopropyl alcohol (rubbing alcohol).

Spectrophotometry and Gel Electrophoresis can be used to determine the quality of DNA. A high-quality DNA sample should have an absorbance ratio of between 220 nm to 280 nm. 1.8. A low ratio http://www.mpsciences.com/2021/04/01/types-of-science-products-available/ could be a sign of an issue with the protein binding steps, or a salt carryover from wash or binding buffers.