Why we use EtBr 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.
Why ethidium bromide is used during agarose gel electrophoresis of DNA fragments?
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 DNAand illumination with a UV light source, theDNA banding pattern can be visualized.
How does EtBr bind to RNA?
Ethidium binds between the stacked bases in DNA. Being a large planar molecule it binds by forming close van der Waals contacts between the stacked bases of a single DNA (or RNA) strand as well as between the bases of the complementary strand.
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 much EtBr should be added to agarose gel?
(Optional) Add ethidium bromide (EtBr) to a final concentration of approximately 0.2-0.5 μg/mL (usually about 2-3 μl of lab stock solution per 100 mL gel). EtBr binds to the DNA and allows you to visualize the DNA under ultraviolet (UV) light. CAUTION: EtBr is a known mutagen.
Can we use EtBr for RNA?
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 is a DNA intercalator, inserting itself between the base pairs in the double helix.
How do you use EtBr?
Prepare enough 0.5µg/ml EtBr in water or buffer to completely submerge the gel. This solution is stable for 1-2 months at room temperature in the dark. After the run submerge the gel in the staining solution for 15-30 minutes (depending upon gel thickness).
What is the function of EtBr in agarose gel?
The EtBr works as a color agent that gives color to DNA. EtBr works as a separating agent in agarose gel electrophoresis. EtBr intercalates between DNA base pairs and emits fluorescence under UV light. By using a standard orange filter, the orange-colored DNA can be seen.