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Analysis of Common Components and Principles of Magnetic Bead Extraction Reagents
The magnetic bead method of nucleic acid purification technology uses nano-scale magnetic beads. The surface of the magnetic beads is labeled with a functional group that can react with nucleic acid by adsorption. Magnetic Silica Particle means that the surface of magnetic beads is wrapped with a layer of silicon material to adsorb nucleic acids. Its purification principle is based on the purification method of glass milk. So what are the common ingredients in the reagents using magnetic bead extraction, and what is the principle? The editor of Desheng lists some common ingredients in turn, SDS (sodium dodecyl sulfate)-properties:
1. Anionic surfactant: slightly toxic to human body;
2. Excellent foaming agent: widely used in washing powder, toothpaste, shampoo, fire extinguisher;
3. Easily soluble in hot water: When the water temperature is higher than 8°C (≈), the solubility of SDS increases sharply with the increase of water temperature. When the water temperature rises to 70°C (≈), the solubility increase is not obvious.
SDS (Sodium Dodecyl Sulfate)-Function: It can break the intra- and intermolecular hydrogen bonds, unfold the molecule, destroy the secondary and tertiary structure of the protein molecule, and denature the protein. Under high temperature, it can destroy the binding of protein and nucleic acid and promote the release of nucleic acid.
Desheng TRIS buffer
Protein K (Protein K): It is a powerful proteolytic enzyme isolated from Candida albicans with high specific activity and is a key reagent for DNA extraction. In DNA extraction, the main function is to enzymatically decompose the histone bound to nucleic acid, so that the DNA is free in the solution.
Polyethylene glycol octyl phenyl ether (Triton X-100-Triton): Triton X-100 is a non-ionic surfactant, which does not dissociate in water, has high stability in solution, and is not easy to Affected by strong electrolyte inorganic salts, it can combine with lipids such as phospholipids in biological membranes to form soluble complexes; the hydrophobic end can also combine with the hydrophobic regions of membrane proteins to form complexes and dissolve in solution.
Guanidine Isothiocyanate (GITC): A strong protein denaturant, which can quickly dissolve protein and cause cell structure to break. Nucleoprotein is quickly separated from nucleic acid due to the destruction of its secondary structure. GITC is also the main component of the classic RNA extraction reagent TRIZOL.
Ethylenediaminetetraacetic acid (EDTA): is an excellent calcium and magnesium ion chelating agent. Inhibit DNA degradation by deoxyribonuclease (DNase requires a certain amount of metal ions as prosthetic groups).
Tris-Hcl: The buffer formed by mixing Tris solution and hydrochloric acid, provides a more suitable environment for the lysis solution and nucleic acid. Tris developed and produced by Desheng is a very good choice.
Almost all classic lysates contain detergents (such as SDS, Triton X-100, NP-40, Tween 20, etc.) and salts (such as Tris, EDTA, NaCl, etc.). The role of salt, in addition to providing a suitable lysis environment (such as Tris), also includes inhibiting the nuclease in the sample from destroying nucleic acids during the lysis process (such as EDTA), maintaining the stability of the nucleic acid structure (such as NaCl), etc.
Among the above reagents, Desheng currently sells ethylenediaminetetraacetic acid, TRIS, and TRIS-HCL. The market feedback of these products is very good. In addition, Desheng’s inactivated virus transport medium contains lysate. , It can quickly crack the virus and release nucleic acid, and the sales of the product have been very good.