The Basics of DNA Purification

Before conducting any PCR reaction as well as cloning experiments or DNA sequencing, it’s essential to have a high-quality DNA that is free of contaminants, such as protein, debris and RNA. The process of purifying DNA is known as DNA isolation and is one of the most vital steps in molecular biology. In this article, you’ll discover the fundamentals of DNA purification as well as how to optimize your DNA extraction strategies to get better results.

The initial 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 and so that it will easily separate from the other components of the sample. Once the DNA has been placed in an alkaline and water solution, it is treated with chaotropic and detergents to remove cell membranes and nuclei to release DNA (cell lysis). RNase can also be added to eliminate any contamination of RNA from the sample.

DNA is then separated from other cellular components such as proteins and lipids, using organic solvents like chloroform and phenol. After the DNA is separated from lipids or proteins it is then precipitated using alcohol or ruby alcohol.

Spectrophotometry and Gel Electrophoresis can be used to determine the purity of DNA. A high-quality DNA sample should have an absorbance between 260 and 280 nm of 1.8. A low ratio may indicate that there is a problem with the protein binding processes, or salt carryover from the wash or binding buffers.

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