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Ethidium bromide (or homidium bromide, [2] chloride salt homidium chloride) [3] [4] is an intercalating agent commonly used as a fluorescent tag (nucleic acid stain) in molecular biology laboratories for techniques such as agarose gel electrophoresis.
The most commonly used dye in agarose gel gel electrophoresis of DNA and RNA, dating as far back as the 1970s, is ethidium bromide (2,7-diamino-10-ethyl-9-phenylphenanthridiniumbromide). [citation needed] Ethidium Bromide (EtBr) is an orange-colored fluorescent intercalating
The illuminator apparatus mostly also contains imaging apparatus that takes an image of the gel, after illumination with UV radiation. The ethidium bromide fluoresces reddish-orange in the presence of DNA, since it has intercalated with the DNA. The DNA band can also be cut out of the gel, and can then be dissolved to retrieve the purified DNA.
Acridine orange can be used in conjunction with ethidium bromide or propidium iodide to differentiate between viable, apoptotic, and necrotic cells. Additionally, acridine orange may be used on blood samples causing bacterial DNA to fluoresce, aiding in the clinical diagnosis of bacterial infections, such as meningitis. [3]
Agarose gel with UV illumination: DNA stained with ethidium bromide appears as glowing orange bands. DNA as well as RNA are normally visualized by staining with ethidium bromide , which intercalates into the major grooves of the DNA and fluoresces under UV light.
EtBr allows one to easily visualize DNA or RNA on a gel as EtBr fluoresces an orange color under UV light. [23] Ethidium bromide binds nucleic acid chains through the process of Intercalation. [3] While Ethidium bromide is a popular stain it is important to exercise caution when using EtBr as it is a known carcinogen.
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Consequently, ethidium bromide is often used as a marker for apoptosis in cells populations and to locate bands of DNA in gel electrophoresis. The stain may also be used in conjunction with acridine orange (AO) in viable cell counting. This EB/AO combined stain causes live cells to fluoresce green whilst apoptotic cells retain the distinctive ...