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# Experiment #9: Nucleic Acids and RNA/DNA Isolation and Analysis ## Introduction Nucleic acids are present in all living cells and play a crucial role in cellular organization, function, cell division, reproduction, and transmission of hereditary factors. They are involved in protein synthesis a...

# Experiment #9: Nucleic Acids and RNA/DNA Isolation and Analysis ## Introduction Nucleic acids are present in all living cells and play a crucial role in cellular organization, function, cell division, reproduction, and transmission of hereditary factors. They are involved in protein synthesis and act as co-factors/co-enzymes. DNA (deoxyribonucleic acid) and RNA (ribonucleic acid) are two naturally occurring classes of nucleic acids. This experiment focuses on isolating, quantifying, and profiling RNA from yeast, and examining its component properties. Nucleic acids are slightly soluble in cold water, insoluble in alcohol, but readily dissolve in weak alkali metal salts. They precipitate from an alkaline solution when acid is added. ## RNA Preparation from Yeast In isolating RNA from yeast, heating with 10% sodium hydroxide (NaOH) helps to extract the nucleic acid and inactivates nucleases. The RNA is separated from associated proteins and other interfering substances through acid extraction (pH 4-5). Further purification involves treatment with concentrated hydrochloric acid (HCl) and repeated washing with 95% alcohol and other organic solvents to remove any remaining impurities. ## Complete Hydrolysis of RNA Nucleotides Complete hydrolysis of RNA nucleotides results in a mixture of sugar (ribose), purine (adenine and guanine), pyrimidine (uracil and cytosine) bases, and phosphate groups. ## Objectives By the end of the experiment, students should be able to: 1. Prepare RNA from yeast. 2. Analyze the properties of the prepared RNA. 3. Describe the steps involved in DNA isolation and profiling. 4. Appreciate the importance of nucleic acids in daily life. ## Materials ### A. Apparatus * 2 Porcelain chips * 1 Hot plate * 1 Funnel holder * 1 Water bath ### B. Chemicals * For RNA preparation from yeast: * 20 mL 10% NaOH * 1 mL concentrated HCl * 10 g yeast * 35 mL 95% ethyl alcohol