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It is vital to have high-quality DNA that is free of contaminants such as protein, debris and RNA prior to carrying out a PCR or cloning procedure, or DNA sequencing. Purifying DNA is also known as DNA Isolation, and is a vital step in molecular biology. In this article, you’ll learn the fundamentals of DNA purification and how you can optimize your DNA extraction strategies for greater results.

The initial step of the DNA purification process is to make a solution that consists of the mixture of alkaline buffer and water. This buffer makes DNA soluble, and it is able to be separated from other components of the sample. After the DNA is placed in an alkaline and water solution, it is treated with chaotropic and detergents to destroy cell membranes and nuclei and release the DNA (cell lysis). RNase may also be added to remove any contaminants in the RNA sample.

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

Gel electrophoresis and spectrophotometers can be used to determine the purity of DNA. A good quality DNA sample must have a ratio of absorbance between 260 and 282 nm. 1.8. A low ratio could signal an issue with the protein binding processes, or salt carryover from the wash or binding buffers.

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