How does ethidium bromide work as a DNA stain?
The most commonly used stain for detecting DNA/RNA is ethidium bromide. Ethidium bromide is a DNA interchelator, inserting itself into the spaces between the base pairs of the double helix. Ethidium bromide possesses UV absorbance maxima at 300 and 360 nm. Ethidium bromide is a sensitive, easy stain for DNA.
Is ethidium bromide used to detect DNA?
Ethidium Bromide (EtBr) Dye for DNA and RNA Detection Ethidium bromide is the most commonly used dye for DNA and RNA detection in gels. Ethidium bromide has UV absorbance maxima at 300 and 360 nm, and an emission maximum at 590 nm. The detection limit of DNA bound to ethidium bromide is 0.5 to 5.0 ng/band.
What color does ethidium bromide stain?
Ethidium bromide (2,7-diamino-10-ethyl-9-phenylphenanthridinium bromide) is used as a nucleic acid stain which fluoresces in the presence of ultraviolet (UV) light. It is commonly sold in a powder form which is soluble in water. The powder is dark red or purple in color.
How is ethidium bromide contamination detected?
Some facilities use a handheld UV lamp to check for residual ethidium bromide contamination following spill cleanup. A reddish-orange fluorescence can be detected under both “long” and “short” UV wavelengths.
What does ethidium bromide do?
Ethidium bromide is the most commonly used dye for DNA and RNA detection in gels. Ethidium bromide is a DNA intercalator, inserting itself between the base pairs in the double helix.
How does DNA stain work?
The most commonly used fluorescent DNA stain is Ethidium Bromide (EtBr). Individual EtBr molecules can squeeze between neighboring base pairs in a DNA double helix in a process known as “intercalation”. When excited with UV light, any EtBr intercalated into the DNA fluoresces and produces a bright orange light.
What is ethidium bromide and its use?
Ethidium bromide is commonly used as a non-radioactive marker for identifying and visualizing nucleic acid bands in electrophoresis. It fluoresces readily with a reddish-brown color when exposed to ultraviolet. light, intensifying almost 20-fold after binding to DNA.
Why is ethidium bromide not used in DNA fingerprinting?
Ethidium bromide was reasoned to cause severe health risks by acting as a mutagen because it intercalates double-stranded DNA (i.e. inserts itself between the strands), deforming the DNA, but that hypothesis has been repeatedly disproved by evidence.
How does ethidium bromide stain RNA?
Ethidium bromide is the most commonly used dye for DNA and RNA detection in gels. Ethidium bromide is a DNA intercalator, inserting itself between the base pairs in the double helix. Ethidium bromide has UV absorbance maxima at 300 and 360 nm, and an emission maximum at 590 nm.
Does ethidium bromide stain clothes?
I shouldn’t worry about it. The EtBr will have happily washed down the sink when you washed your trousers, so the rest of your clothing is fine. Don’t panic about it. Any traces will be so low as to be at less-than-homeopathic strength.
How is ethidium bromide contamination removed?
The decontamination solution is prepared by adding 20mL of 50% hypophosphorous acid to a solution of 2g of sodium nitrite in 300mL of water. Scrub the contaminated surface or equipment with a paper towel soaked in the freshly prepared decontamination solution.
How do you decontaminate spillage of EtBr in the lab?
Spill Clean-up: Use ultraviolet light to locate the spill. Then prepare decontamination solution by mixing of 4.2 grams of sodium nitrite and 20 mL of hypophosphorous acid (50%) in 300 mL of water. To decontaminate the spill area, wash it with a paper towel soaked in the decontamination solution.
How does ethidium bromide affect the mobility of DNA?
The effect of ethidium bromide on mobility of DNA fragments in agarose gel electrophoresis Ethidium Bromide (EtBr) is sometimes added to running buffer during the separation of DNA fragments by agarose gel electrophoresis.
How is ethidium bromide used in gel electrophoresis?
Ethidium Bromide (EtBr) is sometimes added to running buffer during the separation of DNA fragments by agarose gel electrophoresis. It is used because upon binding of the molecule to the DNA and illumination with a UV light source, the DNA banding pattern can be visualized.
How is ethidium bromide used in veterinary medicine?
Applications. Ethidium bromide has also been used extensively to reduce mitochondrial DNA copy number in proliferating cells. Effect of EtBr on mitochondrial DNA is used in veterinary medicine to treat trypanosomosis in cattle, as EtBr binds molecules of kinetoplastid DNA and changes their conformation to Z-DNA form.
Why is ethidium bromide used as an intercalating agent?
Ethidium bromide is known as an intercalating agent. The flat structure of ethidium bromide allows it to intercalate, or insert, between nitrogenous bases of a DNA molecule. This interaction is especially useful because of ethidium bromide’s second characteristic.