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Ethidium Bromide in agarose gel electrophoresis
The principle of Ethidium Bromide in DNA Staining and UV detection:
* Ethidium bromide intercalates between DNA base pairs
* UV light at 312 nm causes the ethidium bromide /DNA combination to fluoresce
Basic protocol of 1 mini gel preparation with ethidium bromide:
* Melt agarose powder in 50ml gel solution without ethidium bromide.
* Swirl solution to cool down a little, then mix 2ul of 10ml/ml of ethidium bromide to it.
* Pour the ethidium bromide containing solution to gel tray to form the gel.
Advanced method of gel casting with ethidium bromide:
1. Prepare 500ml gel solutions in microwave and then add ethidium bromide.
2. Pour 10 gels at once and stack gel tray together for gel storage to a month.
3. Take one gel to run 6 minute high speed gel anytime when needed.
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Safety of ethidium bromide:
Possible routes of
ethidium bromide exposure:
Inhalation - Operations capable of generating ethidium bromide dust, or aerosols of ethidium bromide solutions provide potential for inhalation.
Absorption - May be absorbed through skin.
Effects of ethidium bromide overexposure: Acute toxic effects from exposure to ethidium bromide have not been thoroughly investigated. Ethidium bromide is irritating to the eyes, skin, mucous membranes, and upper respiratory tract.
Physical Hazards Associated with ethidium bromide:
Corrosivity: Irritant
Flammability: Ethidium bromide does not pose a flammability hazard.
Storage and Handling Requirements for ethidium bromide:
Store ethidium bromide in a cool, dry place away from strong oxidizing agents. Keep containers tightly closed. Use with adequate ventilation.
Special Precautions: When handling solid, avoid raising dust.
Engineering Controls:
Use only inside a chemical fume hood with adequate ventilation.
Safety shower and eye wash should be easily accessible where ethidium bromide is used.
Personal Protective Equipment.
Wear chemical safety goggles when using ethidium bromide.
Avoid skin contact, ethidium bromide may be absorbed through the skin. Use rubber gloves.
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