It is important to have a high-quality DNA that is free of contamination such as proteins, debris and RNA prior to carrying out the PCR, cloning, or DNA sequencing. Purifying DNA is also referred as DNA Isolation and is a crucial step in molecular biology. This article will click this link now guide you through the basics of DNA extraction and how to optimize it to achieve better results.
The first step of the DNA purification process is to prepare a solution that contains a mixture of water and an alkaline buffer. This buffer makes DNA soluble and it is easily separated from other components in the sample. Once the DNA is in an alkaline and water solution, it’s treated with detergents or chaotropic salts to destroy cell membranes and nuclei to release DNA (cell lysis). RNase may also be added to eliminate any contamination of RNA from the sample.
DNA is separated from other cellular components like proteins and lipids, using organic solvents like phenol and chloroform. Once the DNA has been removed from 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 should have an absorbance at 260 nm to 282 nm. 1.8. A low ratio may indicate problems with the protein binding steps, or a salt carryover from wash or bind buffers.