Before performing the PCR reaction or cloning test or DNA sequencing, it’s essential to have a high-quality DNA that is free of contaminations such as debris, proteins and RNA. Purifying DNA is also referred as DNA isolation and is a crucial step in molecular biology. In this article, you’ll be taught the fundamentals of DNA purification and how to optimize your DNA extraction strategies for greater results.
The first step in the DNA purification process is to prepare a solution that contains an amalgamation of water and alkaline buffer. This buffer makes the DNA soluble, so that it can easily be separated from other components of the sample. After the DNA is placed in a water and alkaline solution, it is then treated by chaotropic salts or detergents to destroy cell membranes and nuclei to release DNA (cell lysis). RNase can be added to the sample to eliminate any contaminating DNA.
The DNA is separated by organic solvents such phenol or chloroform from other components of the cell like proteins and fats. After the DNA has been removed from the proteins or lipids it is then precipitated using alcohol or ruby alcohol.
The purity of the DNA can then be verified using spectrophotometry or gel electrophoresis. A high-quality DNA sample should have an absorbance range of 260 nm and 280 nm.
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