What are the chemicals used in DNA extraction and their functions?

What are the chemicals used in DNA extraction and their functions?

My ultimate Guide for DNA extraction

DNA extraction step	Chemical
Digestion of protein	CTAB, SDS, phenol, chloroform, Nonidet P40, different chaotropic, urea, guanidium isothiocyanate, guanidium thiocyanate, N-Lauroyl sarcosine
Precipitation of DNA	Isopropanol, ethanol, methanol, NaCl, sodium acetate
Washing of DNA	Any alcohol

What is the specific purpose of detergent used for DNA extraction?

Detergent contains sodium laurel sulfate, which cleans dishes by removing fats and proteins. It acts the same way in the DNA extraction protocol, pulling apart the lipids and proteins that make up the membranes surrounding the cell and nucleus. Once these membranes are broken apart, the DNA is released from the cell.

What is the function of liquid nitrogen in DNA extraction?

Liquid nitrogen is used as it has a very low temperature of -176° C which help to pulverize the hard substance of plant and animal tissue to turn into dust. It also help in deactivating the DNAase to digest the DNA .

What is used in DNA extraction?

DNA purification from detergents, proteins, salts and reagents used during the cell lysis step. The most commonly used procedures are: Ethanol precipitation usually by ice-cold ethanol or isopropanol. Since DNA is insoluble in these alcohols, it will aggregate together, giving a pellet upon centrifugation.

What is the function of chloroform in DNA extraction?

The main function of chloroform is to protect genomic DNA during a catastrophe. Chloroform increases the efficiency of phenol to denature the protein. Here, chloroform allows proper separation of the organic phase and aqueous phase and keeps DNA protected into the aqueous phase.

Which chemical is used to precipitate DNA during procedure of DNA isolation?

Isopropanol/ethanol Isopropanol/ethanol is used in precipitation of DNA, which breaks the hydration shell. Isopropanol is a good choice for precipitation of DNA.

Why is ethanol used in the DNA extraction process?

The initial role of the ethanol and monovalent cations is to remove the solvation shell surrounding the DNA and permitting the precipitation of the DNA in pellet form. The ethanol also serves to promote the aggregation of the DNA. This permits the solubilisation of the salts whilst minimising the solubility of the DNA.

Why is chilled ethanol used for DNA extraction?

Using ice-cold ethanol and ice-coldwater increases the yield of DNA. Low temperatures protect the DNA by slowing down the activity of enzymes that could break it apart. DNases in the cytoplasm would destroy the DNAof viruses entering the cell. Cold ethanol helps the DNA to precipitate more quickly.

What is the role of phenol in DNA extraction?

Extraction of DNA containing samples with acidic phenol results in the denaturation of the DNA, and once denatured, the DNA partitions to the organic phase. This is a key feature of many RNA purification protocols, which is one of the reasons acidic buffer-saturated phenol is used.

Why Isopropanol is used in DNA extraction?

The overall function of salt and ethanol/ isopropanol is to precipitate DNA from the solution. The salts neutralize the negative charge of the negatively charged phosphate in DNA and the isopropanol /ethanol removes the hydration shell of H2O molecules around the phosphate.

What chemical is used to stain the DNA?

Ethidium bromide
Ethidium bromide (EtBr) or 3,8-Diamino-5-ethyl-6-phenylphenanthridinium is commonly used as a non-radioactive marker to stain DNA in order to identify and visualize nucleic acid bands in electrophoresis and other gel-based methods of nucleic acid separation.

Which chemical is used to precipitate DNA during the procedure of DNA isolation?

Isopropanol/ethanol Alcohol is used to precipitate the DNA out of the extraction solution, so we can wash all those salts and chemicals away and then dissolve it in our final solvent—usually water or some variant of Tris-EDTA solution.