Why is EDTA used in DNA extraction?

EDTA (ethylenediaminetetraacetic acid) is a chelating agent that binds divalent metal ions such as calcium and magnesium. EDTA can be used to prevent degradation of DNA and RNA and to inactivate nucleases that require metal ions. EDTA can also be used to inactivate metal ion-requiring enzymes.

What is the purpose of EDTA?

A chemical that binds certain metal ions, such as calcium, magnesium, lead, and iron. It is used in medicine to prevent blood samples from clotting and to remove calcium and lead from the body. It is also used to keep bacteria from forming a biofilm (thin layer stuck to a surface).

Why is EDTA used in gel electrophoresis?

In agarose gel electrophoresis, EDTA is added in buffer for chelating the magnesium ions which are cofactors for DNA nucleases. Hence, activity of DNA nucleases that may be present is inhibited, and DNA is protected from degrading by DNA nucleases.

Why EDTA is used in most of the buffer preparation?

EDTA is a chelator of divalent cations, particularly of magnesium (Mg2+). As these ions are necessary co-factors for many enzymes, including contaminant nucleases, the role of the EDTA is to protect the nucleic acids against enzymatic degradation.

What is the role of isopropanol in DNA extraction?

Since DNA is insoluble in ethanol and isopropanol, the addition of alcohol, followed by centrifugation, will cause the DNA proteins to come out of the solution. In addition, isopropanol is often used for precipitating DNA from large volumes as less alcohol is used (see protocols below).

What is the structure of EDTA in DNA extraction?

The structure of EDTA is shown in the figure below. The EDTA works as a chelating agent in the DNA extraction. It chelates the metal ion present into the enzymes and as we all know that the metal ions are the cofactor which increases the activity of the enzyme.

What is the chemical formula of EDTA?

The chemical formula of EDTA is C10H16N2O8. The structure of EDTA is shown in the figure below. The EDTA works as a chelating agent in the DNA extraction. It chelates the metal ion present into the enzymes and as we all know that the metal ions are the cofactor which increases the activity of the enzyme.

What is EDTA chelation and how does it work?

EDTA is responsible for chelation of divalent ions. It stops the action of DNases found in cytoplasm of cells. For DNA extraction, cells and nucleus need to be disrupted. Therefore, DNA comes in contact with DNases present in the cytoplasm. These DNases, DNA cutting enzymes, can destroy the genomic DNA and reduce the yield of gDNA considerably.

Why is EDTA used to deactivate nuclease enzymes?

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).