What are the methods of staining DNA using ethidium bromide?

What are the methods of staining DNA using ethidium bromide?

Method I – Including Ethidium Bromide in the Gel and Buffer

1. Dissolve agarose in buffer as per the standard protocol for preparing an agarose gel.
2. Allow gel to cool to 60-70°C.
3. Add EtBr to 0.5 µg/ml final concentration. (Stocks are generally 10 mg/ml, and require 5µl stock/100ml gel).
4. Pour gel and allow to set as usual.

What does ethidium bromide react with?

Ethidium bromide binds DNA by intercalating between base pairs, which causes the DNA helix to partially unwind. Deoxyribonucleic acid bands in gels stained with ethidium are fluorescent on exposure to ultraviolet light.

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 does ethidium bromide intercalation with DNA?

Ethidium bromide is known as an intercalate agent as it inserts itself between the stacked bases of DNA. The ring structure of ethidium bromide resembles the rings of DNA bases and it is also hydrophobic. Ethidium bromide is fluorescent and is visible under UV light. This allows DNA to be visualized.

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.

Which techniques ethidium bromide is used?

Ethidium bromide (or homidium bromide, chloride salt homidium chloride) is an intercalating agent commonly used as a fluorescent tag (nucleic acid stain) in molecular biology laboratories for techniques such as agarose gel electrophoresis.

What is ethidium bromide EtBr )? What is the function of EtBr in this experiment and what precaution that you have to take when you are handling with EtBr?

Ethidium bromide (EtBr) is commonly used as a non-radioactive marker for identifying and visualizing nucleic acid bands in electrophoresis and in other methods of nucleic acid separation.

How do you make EtBr stain?

Ethidium Bromide Solution Preparation and Recipe

1. Prepare 800 mL of distilled water in a suitable container.
2. Add 10 g of Ethidium bromide to the solution.
3. Stir on a magnetic stirrer for several hours to ensure that the dye has dissolved.

Why is EtBr fluorescence with DNA?

The reason for Ethidium Bromide’s intense fluorescence after binding with DNA is the hydrophobic environment found between the base pairs. By moving into this environment and away from the solvent, the EtBr cation is forced to shed any water associated molecules.

How does EtBr interact and affect the DNA?

Ethidium Bromide (EtBr) is sometimes added to running buffer during the separation of DNA fragments by agarose gel electrophoresis. The mode of binding of EtBr is intercalation between the base pairs. This binding changes the charge, weight, conformation, and flexibility of the DNA molecule.

What is the function of EtBr?

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.

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.