Summary of nucleic acid separation and purification methods
With the recent rebound of the global epidemic, nucleic acid testing has become the primary way to detect viruses, and it has also deepened everyone’s understanding of nucleic acids. In fact, the separation and with the recent rebound of the global epidemic, nucleic acid testing has become the primary way to detect viruses, and it has also deepened everyone's understanding of nucleic acids. In fact, the separation and purification of nucleic acid is the primary problem that needs to be solved in genetic engineering or protein engineering research. It is the basic step to initiate other downstream activities (such as sequencing, amplification, hybridization, ligation, cloning and biological detection), and is a modern biological detection experiment. The basic means of technology. There are many commonly used methods for DNA extraction. Here we mainly introduce the phenol-chloroform extraction method, the high salt precipitation method, and the silica medium adsorption method.
The phenol-chloroform extraction method uses phenolic reagents as protein denaturants. The sample is first lysed, then extracted (phenol/chloroform) and then precipitated (absolute ethanol). The role of chloroform is to remove excess phenol and promote the separation of the aqueous and organic phases. The extraction method requires multiple centrifugation, and the steps are complicated and easy to cause cross-contamination. In addition, due to the addition of organic solvents such as benzene/chloroform, there will be residues in the final product, which will affect the subsequent downstream applications of genomic DNA.
The high-salt precipitation method removes protein impurities and separates DNA by adding various proteases. This method effectively eliminates the contamination of reagents, and the extracted DNA has a larger yield and higher purity, but the disadvantage is that the process of digesting the protease takes more time.