It is important to have high quality DNA that is free of contaminants such as protein, debris, and RNA before performing a PCR as well as cloning or DNA sequencing. Purifying DNA is also referred to as DNA isolation and is a crucial step in molecular biology. This article will explain the basics of DNA extraction and how to optimize it to achieve better results.
The first step of the DNA purification procedure is to make a solution consisting of an amalgamation of water and alkaline buffer. This buffer makes DNA soluble, which means it is able to be separated from other components in the sample. Once the DNA has been placed in an alkaline and water solution, it is then treated by chaotropic salts or detergents to break down cell membranes and nuclei, and release the DNA (cell lysis). RNase may also be added to eliminate any contamination RNA from the sample.
The DNA is separated by organic solvents such as phenol or chloroform from other components of the cell like fats and proteins. After the DNA is removed from lipids or proteins, it can be precipitated with ethanol or ruby alcohol.
The purity of the DNA sample can be verified using spectrophotometry or gel electrophoresis. A good quality DNA sample should have an absorbance at 260 nm to at least 280 nm.
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