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What is the function of EDTA in DNA extraction?

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2012-08-09 18:11:09
2012-08-09 18:11:09

EDTA is a chelating agent and has great affinity with matel ions and Mg-ion present in DNase as a cofactor and responsible for DNase action that degrade the DNA,here EDTA bind with Mg-ion and nullyfy the action of DNase.

The nuclear envelope normally protects the DNA from digestion by nucleases. Nuclear envelope is the membrane that surrounds the nucleus and prevents the exposure of its contents such as the DNA to the contents of cytoplasm. In the process of DNA extraction, we need to break down the nuclear envelope in order to access the DNA. This would expose the DNA to nucleases and if we don't deactivate these enzymes, they will cut and damage the DNA. Nucleases need divalent cations such as Mg2+ to function. In order to deactivate these enzymes we use EDTA which stands for Ethylenediaminetetraacetic acid to our sample tissue. EDTA has four carboxyl groups ( -COOH). In the alkaline condition of the buffer, EDTA becomes negatively charged. The EDTA ions then form covalent bonds with the divalent cations and prevent them from reacting with nucleases. As a result, the enzymes are deactivated and will no longer cause a threat to the DNA.

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Its purpose is to isolate DNA from a protein mixture.

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it interacts with the lipopolysaccharides found in the outer membrane. by this it is helping the work of EDTA. this will cause cell lysis

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Buffer is used to maintain pH in any process. In DNA extraction EDTA used as a chelating agent at pH8. This ensure the isolation process pure, since chelating agent inactivate the contaminating DNAses.

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TE stands for Tris and EDTA. The Tris buffers the water to prevent acid hydrolysis of the DNA/RNA. The EDTA chelates divalent cations that can assist in the degradation of RNA.


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