It is vital to have high quality DNA that is free of contamination such as debris, protein, and RNA before performing the PCR or cloning procedure, or DNA sequencing. The process of purifying DNA is referred to as DNA isolation. It is one of the most vital steps in molecular biology. This article will teach you the fundamentals of DNA extraction and how to improve it for better results.
The initial step in the process of purifying DNA is to prepare a solution which includes a mix of water and an alkaline buffer. This buffer makes the DNA soluble, so that it is able to be separated from the other components of the sample. After the DNA is placed in an alkaline and water solution it is treated with detergents and Chaotropics salts in order to break up the cell membranes and nuclei. This lets the DNA out. RNase is also added to eliminate any contamination RNA from the sample.
The DNA is separated by organic solvents, such as chloroform or phenol from other cellular components including proteins and fats. After the DNA has been separated from proteins and lipids it can be precipitated using ethanol or isopropyl alcohol (rubbing alcohol).
Spectrophotometry and electrophoresis may be used to determine the quality of DNA. A high-quality sample of DNA should have an absorbance range of the 260-nm range and 280-nm range.
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