The Protein Man's Blog | A Discussion of Protein Research

Plasmid isolation is crucial to biology and an essential step in various procedures, including cloning, DNA sequencing, transfection, in vitro translation, blotting, and gene therapy. However, these applications require the isolation of high-purity plasmid DNA. Whereas genomic DNA extraction is simple and straightforward, plasmid DNA extraction can be more complicated.

A DNA Polymerase is a vital biological enzyme that is present in DNA replication. In the process, DNA copies into two daughter DNA molecules and synthesizes a new DNA strand from the existing strand by adding dNTPs to the growing DNA.

Various types of commercial-grade detergents contain elevated levels of sulfhydryl oxidizing agents, peroxides, salts, and carbonyl compounds. The proteins that are isolated by these detergents are highly susceptible to contaminating peroxides and carbonyls. The peroxides oxidize the proteins, and the carbonyl groups will form Schiff’s bases with the proteins that interfere with its structure.

Blocking buffers are used to bind the surface areas that may attach to reagents during the testing process and prevent nonspecific antibodies from interacting with reagents. Blockers can also cause poor reading or inaccurate results.