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. In case of contact with eyes, immediately flush eyes with copious amounts of water for at least 15 minutes.
In case of contact with skin, immediately wash skin with soap and copious amounts of water. If inhaled, remove to fresh air. Laboratory Safety. Search sitewide:.
Ethidium Bromide. Laboratory Safety Biosafety Ethidium Bromide 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. All work with EtBr is confined to RNS to the fume hood, the gel running and pouring benches, and the gel doc areas which are labeled.
Gels are occasionally brought to a designated bench in RNS for slicing. Making an EtBr solution from powder must be done in the RNS fume hood to avoid the potential for an acute inhalation hazard i.
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 minutes. Put gloves and PPE back on, use the designated spatula to remove the gel from the staining tray. Place the gel in to a tray of distilled water may leave for a few minutes if destaining is required.
Use the designated spatula to place the gel on to the light box, and return the spatula to the hood. You must remove gloves to use the computer keyboard when imaging; the keyboard must remain a non-EtBr space for all users. Before decontamination, de-energize any electrical equipment that might need cleaning, and wear the appropriate PPE. This solution will be acidic; wear gloves. Wash the area with a paper towel soaked in the decontamination solution.
Then wash another five times with soaked paper towels using a fresh towel each time. Place all the used towels in a large container and soak them in fresh decontamination solution for at least one hour.
Using a U-V light, check for completeness of decontamination. Adjust the used and acidic decontamination solution and towels to a pH of with sodium bicarbonate. The towels can then be discarded in the trash, and the solution may be rinsed down the sanitary sewer with a strong stream of running water.
These Stock Solutions must either be poured into a suitable screw-top container and disposed of as hazardous waste, or these solutions can be rendered harmless and poured down the sink using one of the three bench-top treatment procedures: AMRESCO Destaining Bag.
Place a Destaining Bag into the solution. Measure out the agarose using wax weighing paper. Put the agarose in an Erlenmeyer flask. Measure out the correct volume of TAE using a graduated cylinder. Swirl the contents of the flask and cover the top with a paper towel. Microwave your gel for however long it takes to melt completely.
Be careful not to burn yourself, because it will be hot. Next add ethidium bromide -- this is a chemical that intercalates DNA and makes it visible under UV light. You might want to wear gloves. The amount of EtBr to add is as follows: of a 0. The casting trays we have have this rubber stuff around the edges that makes a tight seal when you put them in the gel boxes sideways.
They also have notches where you can insert a comb. The size comb to use depends on the width you want your wells to be.
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