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DNA fluorescent probe stains has the advantages of fast, good repeatability, less sample quantity and no radiation, compared with the traditional isotope detection. It has been widely used in automatic DNA sequencing, antibody immunoassay, disease diagnosis, anti-cancer drug analysis and so on. The sensitivity of DNA stain is an important factor affecting the detection results. Therefore, developing a more sensitive stain and avoiding the interference of biological fluorescence background has become a research hotspot.
Characteristics of DNA Stains
If substance B in DNA molecule is replaced with substance A, using the change of fluorescence after the interaction between substance A and DNA molecules to explore the biological function of substance B, this substance a can be used as a fluorescent stain of DNA molecules.
Classification of DNA Stains
DNA stains can be divided into organic fluorescent stains (molecular fluorescent stains), ion fluorescent stains and other types of stains according to the species of substances.
- DNA Organic Fluorescent Stains
Acridine and phenanthridine dyes are the earliest nucleic acid stains. They combine with DNA molecules through embedding or electrostatic attraction to greatly enhance DNA fluorescence. They are small molecule stains with nonspecific sequence. The basic structure of acridine is diphenylpyridine, which is weakly alkaline in aqueous solution. When acridine orange is connected with acidic substances or there are acidic substances, its characteristic spectrum will change to some extent. So far, acridine orange has been used for biological staining. Acridine orange can interact with DNA or RNA. The fluorescence excitation / emission wavelengths of acridine orange combined with double stranded DNA are 502 / 538 nm (water) respectively.
Cyanines, TOTO and YOYO are a class of asymmetric cyanine dyes with high affinity for dsDNA developed by Glazer research group. The dye has no fluorescence in the solution, which reduces the fluorescence background interference in the detection process. Strong fluorescence occurs after binding with dsDNA, and the fluorescence is enhanced about 1000 times.
- DNA Ion Fluorescent Stains
Rare earth ion stain is also a kind of detection reagent for nucleic acid stain. Lanthanides are the most widely used rare earth elements. The interaction between lanthanide ions and nucleic acids can produce the characteristic fluorescence of lanthanide metal ions, which can provide information about the composition of nucleic acids.
- Other Types of DNA Fluorescent Stains
Quantum dot fluorescent stain is a semiconductor nano particle composed of group and group elements. Nucleic acid is connected to the surface of quantum dots to make a stain. Through in vitro hybridization conventional fluorescence analysis, abnormalities and variations of chromosomes can be found, which can replace some organic fluorescent dyes for biological macromolecular detection.