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The Basics of DNA Purification

Before conducting the PCR reaction, cloning experiment or DNA sequencing, it’s essential to have high-quality DNA that is free of contaminants such as debris, proteins and RNA. Purifying DNA is also referred as DNA isolation and is a crucial step in molecular biology. In this article, you’ll learn the fundamentals of DNA purification and how you can optimize your DNA extraction methods for better results.

The first 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 DNA soluble, so it is easily separated from the other components of the sample. Once the DNA has been placed in a water and alkaline solution, it’s then treated with detergents or chaotropic salts to dissolve cell membranes as well as nuclei and release DNA (cell lysis). RNase can be added to the sample to remove any DNA that is contaminating.

The DNA is then separated by organic solvents such chloroform or phenol from other components of the cell including proteins and fats. After the DNA is separated from lipids or proteins, it can be precipitated using alcohol or rubbing alcohol.

The quality of the DNA can be confirmed using spectrophotometry, or gel electrophoresis. A good quality DNA sample should have an absorbance at 260 nm to 282 nm. 1.8. A low ratio could be a sign of that there is a problem with the protein binding processes, or a salt carryover from wash or buffers for binding.

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