It is essential to have a high-quality DNA that is free of contamination such as protein, debris, and RNA before performing the PCR or cloning process, or DNA sequencing. Purifying DNA is also referred as DNA Isolation and is a crucial step in molecular biology. This article will guide you through the fundamentals of DNA extraction and how to improve it for better results.
The first step of the DNA purification process is to prepare a solution that contains an amalgamation of alkaline buffer and water. This buffer makes DNA soluble and it is easily 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 can be added to the sample to eliminate any DNA that is contaminating.
The DNA is then separated by organic solvents like chloroform or phenol from other cellular components like fats and proteins. After the DNA is removed from the proteins and lipids, they can be separated using ethanol or isopropyl alcohol (rubbing alcohol).
The quality of the DNA can then be confirmed using spectrophotometry, or gel electrophoresis. A high-quality DNA sample should have an absorbance range of 220 nm to 280 nm.
http://www.mpsciences.com/2021/04/15/gene-synthesis-and-transcription-processes/
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