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    Home > Biochemistry News > Biotechnology News > EB (Ethidium bromide, ethyl bromide)

    EB (Ethidium bromide, ethyl bromide)

    • Last Update: 2020-10-25
    • Source: Internet
    • Author: User
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    is a highly sensitive fluorescent dye used to observe " > insugar and polyacrylamide gels in >a href"" . Ethyl bromide is excited and emits orange-red signals with a standard 302nm UV transmission instrument and can be photographed using a Polaroid negative or a gel imaging processing system with a CCD image head.


    The most common way to observe DNA in agarose gels is to dye them using fluorescent dye ethyl bromide, which contains a three-ring plane base that can be embedded between the bases where DNA accumulates. Its binding to DNA is almost non-base sequence specific. In a saturated solution with high ion strength, approximately one ethyl bromide molecule is inserted for every 2.5 bases.


    When the dye molecule is inserted, its plane base group interacts vertically with the axis of the spiral and through van der Worrhline with the upper and lower bases. The fixed position of the group and its close proximity to the base cause the dyes that bind to DNA to appear fluorescent, with an increased fluorescence yield compared to the dyes in the free solution. DNA absorbs UV radiation at 254nm and transmits it to the dye, while the combined dye itself absorbs light radiation of 302nm and 366nm.


    in both cases, the absorbed energy is re-launched at 590nm > in the visibleBecause the fluorescence yield of ethyl bromide-DNA complexes is 20-30 times higher than that of dyes that do not bind DNA, DNA stripes as little as 10 ng can be detected when the gel contains free ethyl bromide ingots (0.5ug/ml).


    brominated ethyl ingots can be used to detect single-stranded or double-stranded nucleic acids (DNA or RNA). However, dyes have relatively little affinity for single-stranded nucleic acids, so their fluorescence yield is relatively low. In fact, most fluorescences of single-stranded DNA or RNA dyes are produced by dyes binding to molecules to form shorter in-chain spirals.


    Although the electrophoresis migration rate of line DNA is reduced by about 15% in the presence of the dye, therefore, when the exact size of the DNA fragment (e.g. DNA limit enzyme cut map identification), the gel should be electrophoresis without EB, electrophoresis after the end of the EB staining. After dyeing, there is usually no need to decolor. However, when detecting small amounts of DNA (less than 10ng) fragments, the dyed gel is usually decolored.


    < Ethyl bromide is a strong mutagent, with high carcinogenicity!


    SYBR Green I and EB Ames test results show that EB is prone to organic mutations. (Singer et al., Mutat. Res. 199, 439: 37- 47.)

    .


    < p style - text-align: left;" >


    (1) for solutions with EB content greater than 0.5mg/ml can be treated as follows:


    (1) dilutes EB solution with water To concentrations below 0.5mg/ml;


    (2) adds double the volume of 0.5mol/L KMnO4, mix well, then add an equal amount of 2.5mol/L HCl, mix well, set room temperature for several hours;

    .


    (3) adds a double volume of 2.5mol/L NaOH, mixing and discarding.


    (2) solution with an EB content of less than 0.5mg/ml can be treated as follows:


    (1) Press as follows:



    (2) is filtered with filter paper and the activated carbon is > sealed with filter paper


    CTAB buffer (DNA extract)


    dexane-based triammobromominated ammonium alias: Western Ammonium bromine, bromine trimethylamide, ammonium bromine, ammonium bromide, cetacean melting ammonium bromide; CTAB; CTMAB; HTAB; CTABr; positive soap


    ;cetrimonium Bromide


    -classified quarterly ammonium salt


    code A-15 CAS No. 57-0 9-0 EINECS login number 200-311-3


    molecular C16H33 (CH3) 3NBr


    ."

    molecular weight 364.446


    melting point: 250-237 degrees C water solubility 13 g/L (20 degrees C) purity (content) > 99%


    II, physicochemical properties


    is white or light yellow crystal to powder, irritating odor, soluble in isopropyl alcohol, soluble in water, A large amount of foam is produced during the shock, which can be well matched with cation, non-ion, s surfactant. Excellent penetration, softening, emulsification, anti-static, biodegradability and sterilization and other properties. The chemical stability of this product is good, heat- and light-resistant, pressure-resistant, strong acid-resistant alkali.


    uses natural, synthetic rubber, silicone oil and asphalt emulsions; anti-static, softeners, conditioners for synthetic, natural and fiberglass; conditioners for conditioners; phase transfer catalysts Emulsion foaming agent, surfactant, analysisreagent, polyester silkener, leather greaser, it is also used in flux, solder paste production of surfactant action, strong activity, bright spots, virtual welding, solder full have a certain role.


    HLB value 15.8, melted in hot water, ethanol, chloromethane, easily melted in isopropyl alcohol aqueous solution, slightly soluble in acetone, insoluble in ether. Good surface activity, stability, sterilization and biodegradability. In strong acids and strong alkalis, heat-resistant, light-resistant. It has good matching with non - ion and two - sex ion surfactants and has a good emulsification effect on a variety of greases. It is less irritating to the skin and mucous membranes and has a slight degreasing effect.


    is a cation defleant widely used in wetting, sterilization, anti-static, de-fouling, solubility and so on. Can be used as a fiber-efficient anti-static agent. In industrial and oil field water treatment, it is used as a bactericidal algae agent and a clay peeling agent. Toothpaste is used as an oral fungicide. For shampoo and conditioner, it makes hair easy to comb, smooth and soft.


    is also used to the right as a mineral flotation agent, electroplating liquid corrosion inhibitor, asphalt emulsizer, hard surface cleaner, and organic synthetic phase transfer catalyst. Also has the characteristics of precipitation nucleic acid and acid polysaccharides from low ion strength solution, under these conditions,protein and neutral polysaccharides remain in the solution, in high ion strength solutions, CTAB and proteins and most acid polysaccharides outside the polysaccharide formation complex, but can not precipitate nucleic acid.


    < DNA from a large number of sticky polysaccharide >-producing organisms such as plants and certain Gliland-negative bacteria, including certain strains of E.coli.


    3, important use


    Extraction DNA formula


    CTAB is ammonium 16 alkyl triamyl bromide,

    CTAB extract buffer classic formula </p.

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