DNA Extraction from Strawberries

The first step in working with nucleic acids (DNA and RNA) is to remove the molecules from inside the cell. Different types of cells need to be processed differently in order to release nucleic acids. All cells have a cell membrane, a phospholipid bilayer that separates the internal environment of the cell from the external environment. In eukaryotes, DNA is housed inside the nucleus of the cell which is surrounded by the nuclear membrane, a second double-layered membrane, also composed largely of lipid molecules. When extracting DNA from plant cells, the cell wall must also be considered; some types of plant tissue require grinding or flash-freezing in order to break through the tough cell wall.

Strawberry fruit tissue is an excellent type of tissue to use for demonstration of DNA extraction. First, ripe strawberries are soft and juicy; as the fruit matures, the cells fill up with water and sugar, which make the fruit so delicious! Second, as the strawberry ripens, a series of chemical reactions take place within the cells that lead to the breakdown of long-chain polysaccharides, like cellulose and pectin, that make the cell wall tough. Lastly, cultivated strawberries (Fragaria x ananassa) are the product of a hybridization between multiple strawberry species, and they have an octoploid genome, meaning they have eight sets of chromosomes inside their cells, totalling 56 chromosomes. That translates to lots of molecules of DNA, which increases our yield and makes the DNA easier to visualize

This protocol for extraction of DNA is based largely on the principle of solubility. Solubility refers to the ability of one substance (the solute) to dissolve in another substance (the solvent). Recall that polar substances dissolve easily in polar solvents, but do not dissolve easily in nonpolar solvents, a phenomenon commonly referred to as “like dissolves like.” Water is a polar solvent, and molecules that dissolve easily in water are referred to as hydrophilic. DNA molecules are hydrophilic because the sugar- phosphate backbone of the molecules are highly polar. This means that DNA dissolves in water, so in this experiment, the DNA that is released when the cells are crushed dissolves in the juice/extraction buffer mixture.

Remember, there are two key ingredients in the DNA extraction buffer aside from the water: dish soap and salt. The dish soap acts to break up the lipid molecules that form the cell membrane and the nuclear membrane, which lyses the cell and releases the cellular contents, including DNA. The salt has two functions in the extraction process. It helps to neutralize the charge on the sugar-phosphate backbone, making DNA less soluble in water, and it also makes the DNA molecules stick to each other, so they are easier to visualize and remove from the solution.

Although the chemical reactions described above are all happening when you add the buffer and crush the strawberries, they are not visible with the naked eye. However, the addition of the cold ethanol caused a much more dramatic result! Ethanol is a nonpolar solvent, and when it is added to the solution of strawberry juice and extraction buffer, the DNA precipitates out of the solution. A precipitation reaction is a chemical reaction that causes a solid substance to emerge from a liquid solution. In this experiment, the addition of ethanol to the reaction forces DNA to precipitate out of solution, which we can then spool onto a stirring spoon.

Illustrations of ploidy levels; N=number of chromosomes in one set.Strawberries (Fragaria x ananassa) are octoploid (8N); illustration from Wikimedia Commons

Safety Precautions

Materials

Draw in your lab book what your final product looks like (show the test tube, stirrer, DNA).  How would you describe the texture and appearance of the DNA, including its color?