Plant cells posses rigid cellulose cell wall and a cell membrane.

Breaking the cell, layer by layer

releases the cellular constituents. At first, the plant cell wall has to be damaged. It is
accomplished by grinding the plant tissues and homogenizing in the extraction buffer. The
components of the extraction buffer cause cell lysis.

Tris (Tris-hydroxymethyl aminomethane) in the extraction buffer, buffers the pH of the

cells at 8.0. EDTA (Ethylene diamine tetraacetic acid) deters the DNase activity, chelates the
metal ions which are cofactors for them. It also weakens the cell membrane stability. Sodium
chloride also helps in maintaining the osmoticum of the cells. Chloride component enter into the
cells by ionic transport along with water by diffusion. This influx expands the cells, prevents cell
aggregation and weakens the membrane integrity.

Mercaptoethanol cleaves the disulphide bridges of proteins and helps in denaturation of

membrane proteins and cytosolic proteins. SDS is an anionic detergent, which also denatures the
membrane proteins and disrupts the cell membrane. This also results in the break down of the
nuclear envelope and thereby releases the nucleoplasm. SDS also helps in inhibition of
nucleases.
DNA in the nucleoplasm is neutralized by Potassium acetate. K+ ions bind with negative
phosphate backbone of the DNA and shield them. This favors DNA precipitation from ethanol in
cold condition. Upon centrifugation, the neutralized DNA with impurities goes into the
solution this time. When alcohol is added in cold condition to the solution, DNA strands come
close together and coalesce. But other impurities are soluble in ethanol and are removed in the
supernatant. DNA precipitate is settled down as a pellet by centrifugation, purified by 70 %
ethanol wash, hydrated in TE buffer and analyzed on agarose gel.