Extractor for ethidium bromide decontamination includes. Ethidium bromide laboratory spill and waste handling. The toxic effects of etbr may be experienced if swallowed, inhaled, or absorbed through the skin. Concentrating the toxic dye into a small bag allows the user to dispose of it in the solid waste instead of as a large amount of liquid waste, potentially saving a lot of money in toxic waste disposal fees. It has a role as an intercalator and a fluorochrome. To determine the amount of bound ethidium the following expressions below is used, given that the information of d o and e o are known. The cd of dna ethidium bromide complexes in the 290360 nm region is characterized, especially at high salt and at high ethylene glycol content, by positive and negative. Therefore, it is commonly used as nucleic acid fluorescent tag in various techniques of the life science field. Measuring binding constant of ethidium bromide etbr to dna. Ethidium bromide in water, tbe buffer, mops buffer, and cesium chloride solution may be completely degraded by reaction with sodium nitrite and hypophosphorous acid.
This compound acts as an intercalating agent to stain nucleic acids by marked florescent orange during exposure to uv light. Ethidium bromide solid and stock solutions stock solutions. Ethidium bromide definition is a fluorescent mutagenic biological dye c21h20brn3 that is used especially to stain nucleic acids. Decontamination of ethidium bromide protocol online. Apr 18, 2016 ethidium bromide is found in pretty much every molecular biology lab around. The material fluoresces a redorange color under ultraviolet light, with increased fluorescence when the material is bound to doublestranded dna. Ethidium bromide solution 10 mgml catalog or product number. Ethidium bromide is a very common fluorescent intercalating agent used for visualization of nucleic acids.
Please follow the instructions listed in this fact sheet when handling etbr. Apr 04, 2011 the maximum amount of ethidium bromide that can bind to dna is one molecule per base pair, so this is generally a good place to start in terms of ethidium concentration in the digest reaction. Please contact ehrs at 2157072520 for additional disposal guidance information. In any case you will want to avoid brushing and dusting off the contaminated surface to reduce the formation of ethidium bromide dust. Each unit processes a liter of buffer in less than a minute, removing 99% of etbr and maintains this efficiency for up to 10 liters initial etbr approx. Sansone environmental control and research program, program resources, inc. Ethidium bromide ethidium bromide is an intercalating agent which resembles a dna base pair. A safer alternative is to purchase ethidium bromide in solution. There is no evidence supporting or refuting carcinogenicity in humans. The crystal or powder form is odorless and appears dark red in color. Ethidium bromide etbr wastes are not regulated by the state of massachusetts or the epa. Researchers are rightly wary of this potential carcinogen. Ethidium bromide enhancement of frameshift mutagenesis.
An activated charcoal filter has been used to remove ethidium bromide eb in water, various buffers, cscl solutions and ethanol. Ethidium bromide etbr safety guidance document number. Soak paper towel in decontamination solution, place on contaminated surface, and scrub. Gloves should be changed before touching other surfaces or clean areas. This translates to 30 gml of ethidium bromide in a 20 l digest containing 1 g of plasmid. I now realize to use nitritehypophosphite decontamination procedures. Do not dispose of ethidium bromide as biohazardous waste. The extractor for ethidium bromide decontamination is a selfcontained activated carbon extraction device for removing mutagenic ethidium bromide etbr from electrophoresis buffers. It is commonly abbreviated as etbr, which is also an abbreviation for bromoethane. It fluorescents a redorange color under ultraviolet uv light and with increased fluorescence when bound to doublestranded dna. A logbook entry has to be generated for every disposal entering labnumber. Method for surface decontamination of ethidium bromide. Thats because the compound is used to make dna fluoresce when running gels, and it does that by slipping neatly betw.
Before decontamination, deenergize all electrical equipment and wear the appropriate protective equipment. Etbr is a dark red, crystalline, nonvolatile solid, moderately soluble in water, which fluoresces. It is mutagen or carcinogen, so be careful during the use better use gloves. Aqueous buffer solutions containing ethidium bromide can be released down the drain after decontamination or destruction. It fluoresces a redorange colour under ultraviolet light and with increased fluorescence when bound to doublestranded dna. Ethidium bromide c 21 h 20 brn 3, also known as bromoethane h 3 cch 2 br or simply etbr, is used in the biochemistry by detecting nucleic acids. How to decontaminate the working zone with ethidium bromide hello dear friends.
Ethidium bromide etbr is a mutagen, a suspected carcinogen and teratogen, and at high concentrations is irritating to the eyes, skin, mucous membranes and upper respiratory tract. Ethidium bromide gels, stock solutions, buffers and contaminated debris should be referred to ehs for collection and disposal. Our 5 ml ethidium bromide solution, is supplied in a convenient dropper bottle. Ethidium bromide intercalates inserts itself between nucleic acid base pairs and exhibits fluorescence under ultraviolet light, providing molecular biologists with an excellent stain for visualizing and imaging nucleic acids. Reduce vapor concentrations by covering the surface of a liquid spill with. Ethidium bromide ethidium bromide2,7diamino10ethyl9phenylphenanthridinium bromide etbr remains a widely used dye for the visualization of nucleic acids in agarose gels. Ethidium bromide is the most commonly used dye for staining dna during or after polycrylamide or agarose gel electrophoresis. Ethidium bromide etbr, a dark red, crystalline, nonvolatile powder that is moderately soluble in water, is widely used in research laboratories as a visualizing agent for nucleic acids. Destruction and decontamination of solutions 2 george lunn and eric b. Decontamination of ethidium bromide background ethidium bromide etbr. Wipe the area with a damp cloth after the work with ethidium bromide powder is complete.
Though not a carcinogen, it behaves as a mutagen in the ames salmonella bioassay. Ethidium bromide etbr is commonly used as a marker for identifying and visualizing nucleic. Ethidium bromide is mutagenic and moderately toxic and must be handled with. Ethidium bromide is a wellknown and widely used fluorescent dye in biotechnology research. Ethidium bromide is not a regulated hazardous material, but due to its potential health hazards, precautions should be taken during handling and disposal. Filtering aqueous ethidium bromide waste solutions that are free of other contaminants through a bed of activated charcoal is a relatively simple and effective method for removal of ethidium bromide. Once bound to nucleic acids, the fluorescence of the dye is enhanced by more than 10 times while the excitation maximum is red shifted by 3040 nm and emission is blue. Due to its unique structure, it can easily intercalate into dna strand.
In the case of dna this is usually doublestranded dna from pcrs, restriction digests, etc. Ethidium bromide 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 cd of dnaethidium bromide complexes in the 290360 nm region is characterized, especially at high salt and at high ethylene glycol content, by positive and negative. Ultrapure ethidium bromide is a sensitive fluorescent dye used to detect nucleic acids in agarose gels and caesium chloride cscl gradients. Lunn and sansone method for the decontamination of equipment contaminated with ethidium bromide lunn, george, and sansone, eric b. This mutagenic, teratogenic nucleic acid staining compound should be well decontaminated when cleaned well. Wear appropriate protective eyeglasses or chemical safety goggles as described by oshas eye and face protection regulations in 29 cfr 1910. More recently this wariness has been whipped up into a witch hunt by companies touting safer alternatives and disposal methods. Ethidium bromide etbr is a powerful mutagen and is moderately toxic.
Clean up gross or suspected areas with wet absorbent cloth and water, repeat several times with decontamination solution, repeat several times again with water and absorbent cloth to remove decontamination solution. Ethidium bromide safe handling, disposal, and emergency procedures. Ethidium bromide etbr is commonly used as a nonradioactive marker for identifying and visualizing nucleic acid bands in electrophoresis and in other methods of nucleic acid separation. Since eb can be destroyed by heating at temperatures above 262. Treatment of ethidium bromide solutions with bleach is not permitted. Eb is an intercalating dnabinding dye with little sequence preference. This information will allow us to calculate the binding constant of ethidium toward dna. Ethidium bromide safe handling and disposal procedures. Like its groundbreaking predecessor, this second edition of destruction of hazardous chemicals offers a collection of detailed procedures that can be used to degrade and dispose of a wide variety of hazardous chemicals. They can be also detected with ultraviolet lights, but they are dangerous to the humans eye, so this is where ethidium bromide stain takes part in and makes the availability for the nucleic. Note that studies have shown that treatment with hypochlorite does little to diminish the mutagenic properties of ethidium bromide. Its toxicological properties are not fully determined. Ethidium bromide is strongly mutagenic, causing living cell mutations. Ethidium bromide is found in pretty much every molecular biology lab around.
In all cases studied, ethidium bromide was undetectable in the eluted liquid contamination less than 0,5. Ask most biologists about handling it, and youre get a fearful expression and advice to use gloves, etc. Absorption method for rapid decontamination of solutions. Disposal, decontamination, and destruction procedure. Personal protective equipment ppe to include, at a minimum, lab coat and nitrile gloves should be worn when working with solutions that contain this dye. The book has been expanded and updated to broaden the scope of chemicals treated and to include new and modified procedures and alternatives to the use of some highly toxic.
It is a phenanthridinium intercalator, structurally similar to propidium iodide. Add sufficient water to reduce the concentration of ethidium bromide to 0. Ethidium bromide waste disposal fact sheet environmental. Solutions were excited at 546 nm by a 200w hg lamp filtered by cor ning cs 369 and 496 filters. One kit has the capacity to remove 500 mg of ethidium bromide from solutions 10 mg etbrbag. Etbr may also cause extreme eye and skin irritation. Excitation with light at a wave lenght of 254 nm is absorbed by the dna and transferred to ethidium bromide. Ethidium bromide is commonly used to detect nucleic acids in molecular biology laboratories. Ethidium is the fluorescent compound widely used in experimental cell biology and biochemistry to reveal doublestranded dna and rna.
Ethidiun bromide is being used for nucleic acid staining. The why and how of ethidium bromide assisted partial digests. The activated charcoal decontamination bags concentrate ethidium bromide from used gel staining solutions in a convenient bag. Ethidium azide analogs 3amino8azidoethidium monoazide and ethidium diazide have been developed as photosensitive probes in order to analyze directly the reversible in vivo interactions of.
Ethidium bromide enhancement of frameshift mutagenesis caused by photoactivatable ethidium analogs. Are other fluorescent tags used instead of ethidium. The compound forms fluorescent complexes by intercalation and these compounds are readily visible under ultraviolet uv light. This eliminates the hazards of working with the powdered form. Agarose gel electrophoresis is one of them figure 2, in that etbr is generally used. Wear gloves and a face mask when working with ethidium bromide in powdered form. Ethidium bromide stock solution dissolve 50 mg ethidium bromide in 5 ml double dist. Ethidium bromide etbr is a commonly used stain for the visualization of nucleic acids in agarose gels. Ethidium bromide fluoresces when it is in a hydrophobic environment, such as when its stuck between two base pair stacks in dna water quenches its fluorescence. The powder form is considered an irritant to the upper respiratory track, eyes and skin. Yes, alconox powdered precision cleaner is excellent at emulsifying and decontaminating ethidium bromide on surfaces.
Feb 27, 2020 ethidium bromide is a potent mutagen which has been commonly used as a nucleic acid stain for many years. Ethidium bromide eb is a widely used nucleic acid gel stain although it is also a known potent mutagen. Analytical biochemistry 162, 453458 1987 ethidium bromide. While it is not specifically regulated as a hazardous waste, the mutagenic properties may. Waste stream description waste management procedure. Thermo scientific pierce ethidium bromide is available as a dilute 0. How to decontaminate laboratory equipment contaminated. Typically eb is purchased in powder or solution form and is soluble in water. The powder is used to make a stock solution of 10 mgml, and the working concentration for gelstaining is 1 gml. Some will just use 70% ethanol and restrict this areamachine and assigned it for ethidium bromide stuff forever.
Thats because the compound is used to make dna fluoresce when running gels, and it does that by slipping neatly between the base pairs intercalation, like sliding a card into a deck. Mechanism of ethidium bromide fluorescence enhancement. The hysteria among molecular biologists about our old friend ethidium bromide has long been an irritation to me. Absorption method for rapid decontamination of solutions of. Instructions ethidium bromide disposal 03012006 ethidium. Safe method of use 16 disposal and decontamination of. Personally, i would recommend to wipe all surfaces first with water according to wikipedia the solubility is 40 g l until clean using paper towels and.
Singlestranded rna can also be detected, since it usually folds back onto itself and thus provides local base pairing for the dye to intercalate. I also found this article citing the fact that sunlight can break down etbr but only when a catalyst of iron and titanium dioxide is there to catalyze the reaction. The interaction of the intercalating dye ethidium bromide with several native and synthetic polydeoxyribonucleic acids has been studied by means of circular dichroic spectra. Decontamination of ethidium bromide with biotiums activated charcoal bags compared to resin decontamination bags from another supplier.
Destruction of hazardous chemicals in the laboratory. Ethidium bromide solutions and buffer solutions containing ethidium bromide should be filtered, decontaminated, or destroyed using one of the methods below or collected as chemical waste. Detect as little as little as 1 ng of nucleic acids in agarose gels. To avoid confusion, some laboratories have used the abbreviation ethbr.
Ethidium bromide is typically purchased in powder or solution form and is soluble in water. Scrub five more times with paper towels soaked in water, using fresh towel each time. Use a uv light to survey work surfaces in the laboratory to ensure that the ethidium bromide has been removed. The powder is used to make a stock solution of 10 mgml, and the working concentration for gelstaining is 1. It is sometimes the cause of health and safety concerns for workers charged with using it during the course of their work. Environmental health and radiation safety temple university. Ethidium bromide etbr is a wellknown fluorescent tag figure 1 that is usually applied in molecular biological techniques. The dates for the thorough cleaning must be entered into the log book. To be safe, you can try using sybr safe from molecular probes as an alternative to ethidium bromide. Cleaning of equipment and laboratory surfaces contaminated with etbr. Ethidium bromide definition of ethidium bromide by. Ethidium bromide laboratory spill and waste handling procedures. Each drop contains 25 g of ethidium bromide and is ideal for a 50 ml agarose gel after.
We are a research lab and would like to know if the powder detergent alconox serves for decontaminating ethidium bromide on surfaces. The book has been expanded and updated to broaden the scope of chemicals treated and to include new and modified procedures and alternatives to the use. How to decontaminate the working zone with ethidium bromide. Ethidium bromide is typically in powder or solution form and is soluble in water. Procedure for cleanup of laboratory surfaces contaminated with ethidium bromide. Ethidium bromide is a potent mutagen used as a nucleic acid stain.
Use soap and water mixture detergent solution or 70% ethanol to wipe clean laboratory work surfaces contaminated with ethidium bromide. The filtrate may be poured down the drain the used filters are disposed of similar to gels. Use granular charcoal filtration see appendix d in the lab safety book and chemical hygiene. Procedure for cleanup of laboratory surfaces contaminated. The etbr green bag kit allows rapid and troublefree concentration of ethidium bromide from large volumes of solutions into a small tea bag which is then conveniently disposed along with other solid hazardous wastes. It is available as an aqueous solution containing 10 mgml ultrapure ethidium bromide and highpurity water. Treatment of ethidium bromide solutions with bleach is not permitted method 1 lunn and sansone 1987 1. Solutions were protected from light and either left still or stirred constantly using a magnetic stirbar during decontamination. Ethidium bromide, dropper bottle convenient 5 ml bottle. How to decontaminate laboratory equipment contaminated with etbr. Standards for collection, storage, handling and disposal of human biological material.
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