Experiment:

Extraction and Estimation of DNA/RNA from plant sources

Materials & Apparatus

Any available plant material, mortar and pestle, test tubes, beakers, ethanol,
spool/glass rod, digestive enzymes (cellulase, lipases, protease, ribonucleases
(RNases)/deoxyribonuleases (DNases)

Remember: ribonucleases (RNases) in case of DNA extraction and deoxyribonuleases

(DNases) in case of RNA extraction

Procedure

· Take the available plant material and grind it in the mortar.

· Treat the material with cellulase to break down the cell wall of the plant cells.

· Next, treat it with protease to hydrolyse the peptide bonds of proteins in the plant
material. In other words, the enzyme removes the histone proteins which are
intertwined with the DNA/RNA.

· Dissolve RNA with ribonuclease OR DNA with deoxyribonuclease (WITH

REFERENCE TO PARTICULAR NUCLEIC ACID EXTRACTION)

· Use lipase to dissolve lipids.

· Add chilled ethanol to enable the precipitation of the DNA OR RNA. It essentially
increases concentration.

· Use spooling to extract the precipitated DNA/RNA. Spooling involves winding the
fine threads of DNA/RNA on to a reel.

For quantitating DNA or RNA, readings should be taken at wavelengths of 260nm by

ultraviolet (UV) absorbance spectrophotometry. The amount of UV radiation absorbed by the
solution of DNA or RNA is directly proportional to the amount of DNA or RNA in a sample.