How do intercalating agents cause mutations?
Intercalating agents are another type of chemical mutagen. They tend to be flat, planar molecules like benzo[a]pyrene, a component of wood and tobacco smoke, and induce mutations by inserting between the stacked bases at the center of the DNA helix.
What do intercalating agents do?
Agents that are capable of inserting themselves between the successive bases in DNA, thus kinking, uncoiling or otherwise deforming it and therefore preventing its proper functioning. They are used in the study of DNA.
How does an intercalating agent such as ethidium bromide cause mutations?
As you said EtBr is chemical mutagen act as a intercalating agent. These intercalating agents are flat molecules that can slip between base pairs in the double helix, slightly unwinding the helix and hence increasing the distance between adjacent pairs. This way these molecules cause the mutation.
What are DNA intercalators?
Intercalators are compounds that usually have planar aromatic moieties and can insert themselves between adjacent base pairs in the DNA helix. These types of interactions change the structure of DNA, leading to various types of disorders in the functioning of cells and the cell cycle.
How does ethidium bromide bind to DNA?
Ethidium is capable of forming close van der Walls contacts with the base pairs and that’s why it binds to the hydrophobic interior of the DNA molecule. Ethidium Bromide Binds to DNA. Ethidium binds by inserting itself bewteen the stacked bases in double-stranded DNA.
Why EtBr is 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.
Is ethidium bromide a DNA solution?
Ethidium bromide is commonly used to detect nucleic acids in molecular biology laboratories. In the case of DNA this is usually double-stranded DNA from PCRs, restriction digests, etc. Ethidium bromide is also used during DNA fragment separation by agarose gel electrophoresis.
Is ethidium bromide a mutagen?
Because ethidium bromide can bind with DNA, it is highly toxic as a mutagen. It may potentially cause carcinogenic or teratogenic effects, although no scientific evidence showing either health effect has been found.
Is ethidium bromide flammable?
Not flammable or combustible. Use water spray, alcohol-resistant foam, dry chemical or carbon dioxide. Wear self contained breathing apparatus for fire fighting if necessary. Wear respiratory protection.
What happens if you touch ethidium bromide?
Health and Safety EtBr is a potent mutagen (can cause genetic damage), and moderately toxic after an acute exposure. EtBr can be absorbed through skin, so it is important to avoid any direct contact with the chemical. The powder form is considered an irritant to the upper respiratory tract, eyes, and skin.
Is ethidium bromide a dye?
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.
Is ethidium bromide reactive?
Though there is no evidence of its carcinogenicity or teratogenicity in humans, its mutagenic properties are cause to regard it as a possible carcinogen and reproductive toxin. EB poses no flammability (NFPA rating = 1) or reactivity hazards. It should be stored away from strong oxidizing agents.
Is ethidium bromide still used?
Despites the serious toxicity of EtBr, it is still used in some labs because it is considerably less expensive in comparison to other compounds like SYBR®-based dyes (an asymmetrical cyanine dye used as a nucleic acid stain).
Is ethidium bromide visible under UV?
Ethidium bromide (EtBr) can be incorporated into the gel and running buffer during electrophoresis. As little as 0.05 μg of DNA can be visualized in one band when the gel is exposed to ultraviolet light (Figure 5.4). The EtBr-nucleic acid complex absorbs UV irradiation at about 260 or 300 nm.
Is ethidium bromide cytotoxic?
Cytotoxic chemicals induce damages to the components of DNA and thus results into mutations. for estimating the possible cytotoxic effect of both acridine orange and ethidium bromide respectively. The high mitotic index and lower percentage of abnormality was very much evident in roots treated with diluted mutagens.
What would happen if the gel was run for too long?
If you run gel electrophoresis too long, the sample can run out of the bottom of the gel.
How is ethidium bromide contamination removed?
Charcoal Filtration: Filtering the aqueous EtBr waste solutions (free of other contaminants) through a bed of activated charcoal is a relatively simple and effective method for removal of EtBr. The filtrate may be poured down the drain.
What color is ethidium bromide?
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.
Why is ethidium bromide used in gel electrophoresis class 12?
Ethidium Bromide or EtBr is an intercalating agent. EtBr is commonly used to detect nucleic acids inluding double stranded DNA or single single-stranded RNA. When exposed to UV light, it will fluoresce with an orange colour, intensifying almost 20-fold after binding to DNA.
Why do you need to wear gloves while handling ethidium bromide?
Gloves should be worn when handling ethidium bromide. Disposable nitrile gloves provide adequate protection against accidental hand contact with small quantities of most laboratory chemicals.