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The Basics of DNA Purification

Before conducting a 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 particles, proteins and RNA. Purifying DNA is also known as DNA Isolation and is a vital step in molecular biology. In this article, you will learn the fundamentals of DNA purification as well as how to improve your DNA extraction processes to get better results.

The first step in the process of purifying DNA is to prepare a solution which comprises a mixture of water and an alkaline buffer. This buffer makes DNA soluble and it is able to be separated from other components in the sample. Once the DNA has been placed in an alkaline-water solution, it’s cleaned with detergents as well as chaotropics salts to break up the cell membranes and nuclei. This releases the DNA. RNase can also be added to remove any contaminants in the RNA sample.

DNA is then separated from other cell components like proteins and lipids with the help of organic solvents such as phenol and chloroform. After the DNA has been removed from the proteins and lipids, it can be extracted using ethanol, or isopropyl alcohol (rubbing alcohol).

The quality of the DNA can then be determined by spectrophotometry or gel electrophoresis. A good quality DNA sample will have an absorbance between 260 and 280 nm of 1.8. A low ratio can indicate a problem with the protein binding steps or the transfer of salt from wash or bind buffers.

April 12, 2024

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