{"id":3819,"date":"2016-12-07T14:54:47","date_gmt":"2016-12-07T20:54:47","guid":{"rendered":"https:\/\/wp.stolaf.edu\/chemical-hygiene\/?page_id=3819"},"modified":"2023-08-24T12:56:56","modified_gmt":"2023-08-24T17:56:56","slug":"ethidium-bromide","status":"publish","type":"page","link":"https:\/\/wp.stolaf.edu\/chemical-hygiene\/ethidium-bromide\/","title":{"rendered":"Ethidium Bromide"},"content":{"rendered":"
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Ethidium Bromide (EtBr), a dark red, crystalline, non-volatile powder that is moderately soluble in water, is widely used in research laboratories as a visualizing agent for nucleic acids.  The powder is used to make a stock solution of 10 mg\/mL, and the working concentration for gel-staining is 1 \u00b5g\/mL.  Because EtBr can bind with DNA, it is considered highly toxic as a mutagen.  It may potentially cause carcinogenic or teratogenic effects, although no scientific evidence showing either health effect has been found.  Exposure routes of EtBr are inhalation, ingestion, and skin absorption.  An acute exposure to EtBr causes irritation of the mouth, upper respiratory tract, skin, and eyes.<\/p>\n\n\n\n

Ethidium bromide is not regulated as hazardous waste; however, St. Olaf College policy requires that the following precautions and procedures are taken while using or handling this chemical.<\/p>\n\n\nPersonal Protective Equipment<\/strong><\/span>

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  • Chemical splash-resistant safety\u00a0goggles and gloves MUST be used when handling pure EtBr, preparing ethidium bromide solutions, staining gels, or\u00a0visualizing nucleic acids in gels.<\/li>\n<\/ul>\n<\/div>\n\n\n\n

    <\/p>\n\n\nSpecial Handling Procedures<\/strong><\/span>

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    • All work with\u00a0EtBr<\/span>\u00a0<\/span>is confined to RNS 395 to the fume hood, the gel running and pouring benches, and the gel doc areas (which are labeled). \u00a0Gels are occasionally brought to a designated bench in RNS 375 for slicing.<\/span><\/li>\n
    • Making an EtBr solution from powder must be done in the RNS 395 fume hood to avoid the potential for an\u00a0acute inhalation hazard (i.e., open the container of pure EtBr only inside the RNS 395 fume hood).\u00a0<\/span><\/li>\n
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      <\/span>Before\u00a0you leave the <\/span>EtBr<\/span>\u00a0work area you MUST remove your gloves to prevent <\/span>EtBr<\/span> being transferred to other parts of the lab, or to outside of RNS 395.<\/span><\/p>\n<\/li>\n<\/ul>\n<\/div>\n\n\n\n

      <\/p>\n\n\nPrimary and Secondary Containment; Labeling<\/strong><\/span>

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      • The EtBr\u00a0powder is kept tightly closed and stored in the RNS 395B refrigerator.<\/li>\n
      • The 10 mg\/mL stock solution is to be kept in a tightly closed and labeled container, in a labeled secondary containment tray.<\/li>\n
      • Containers for the 1 \u00b5g\/mL working solutions must be clearly labeled, have lids, and placed in a labeled secondary containment tray.<\/li>\n
      • All primary and secondary containers must be labeled \u201cDesignated for ethidium bromide use only.\u201d<\/li>\n
      • Make sure to include the EtBr concentration on all labels.<\/li>\n<\/ul>\n<\/div>\n\n\n\n

        <\/p>\n\n\nProcedure for Gel Staining<\/strong><\/span>

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        • Wearing all PPE as stated above, place the gel in one of the staining trays containing EtBr in a 1 \u00b5g\/mL concentration.<\/li>\n
        • Close the lid of the tray; you may rinse and remove gloves and leave them at the staining station until gel is ready for viewing (about 10-15 minutes).<\/li>\n
        • Put gloves and PPE back on, use the designated spatula to remove the gel from the staining tray.<\/li>\n
        • Place the gel in to a tray of distilled water (may leave for a few minutes if destaining is required).<\/li>\n
        • Use the designated spatula to place the gel on to the light box, and return the spatula to the hood.<\/li>\n
        • You must remove gloves to use the computer keyboard when imaging; the<\/strong> keyboard must remain a non-EtBr space for all users<\/strong>.<\/li>\n
        • When you are finished viewing and\/or photographing the gel, put it in the designated yellow metal waste container labeled \u201cethidium bromide waste.\u201d<\/li>\n
        • Rinse the Li-Cor EtBr imaging tray with deionized water and then wash with 70% Ethanol and dry with a Kimwipe; the wipes and your gloves should be safe for disposal in the regular trash (turn gloves inside-out).<\/li>\n<\/ul>\n<\/div>\n\n\n\n

          <\/p>\n\n\nSmall Spills; Decontamination of Counters or Equipment<\/strong><\/span>

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