Before conducting the PCR reaction or cloning test or DNA sequencing, it is crucial to have high-quality DNA that is free of contaminations such as protein, debris and RNA. Purifying DNA is also referred to as DNA Isolation and is an essential step in molecular biology. This article will explain the basics of DNA extraction and how to optimize it for better results.
The first step of the DNA purification process is to prepare a solution that includes a mix of water and an alkaline buffer. This buffer makes the DNA more soluble so that it is able to be separated from the other components of the sample. Once the DNA has been placed in an alkaline and water solution, it is then treated with detergents and Chaotropics salts to break down cells’ membranes and nuclei. This lets the DNA out. RNase can also be added to remove any contamination RNA from the sample.
DNA is separated from other cellular components such as proteins and lipids by using organic solvents like phenol and chloroform. After the DNA is separated from lipids or proteins it can be precipitated using ethanol or rubbing alcohol.
Spectrophotometry and gel electrophoresis can be used to determine the quality of DNA. A good quality DNA sample must have a ratio of absorbance of 260 nm up to at least 280 nm.
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