Nucleic Acids and RNA Isolation Quiz

Nucleic Acids and RNA/DNA Isolation and Analysis

• Nucleic acids are vital components in all living cells, playing roles in cell organization, function, division, reproduction, and transmission of hereditary factors
• Nucleic acids function as co-factors and co-enzymes in protein synthesis
• The two main types of nucleic acids are DNA (deoxyribonucleic acid) and RNA (ribonucleic acid)
• The experiment isolates, quantifies, and profiles RNA from yeast, examining its components
• Nucleic acids are moderately soluble in cold water but insoluble in alcohol, mixing instantly in weak alkali metal salts
• RNA isolation involves heating with alkali (10% NaOH) to extract the nucleic acid and inactivate nucleases

RNA Isolation Procedure

• Heating with 10% NaOH extracts RNA and removes water-soluble proteins and nucleases
• Acid extraction at pH 4-5 separates nucleic acid from other interfering substances
• Treatment with 6N acetic acid, followed by repeated washing with 95% alcohol and other organic solvents removes further interfering substances
• This ensures accurate chemical testing

RNA Properties and Objectives

• Complete hydrolysis of RNA yields sugars (ribose), purines (adenine & guanine), pyrimidines (uracil & cytosine), and phosphates.
• Students should be able to prepare RNA from yeast, test its properties, discuss DNA isolation techniques, and appreciate the importance of nucleic acids in daily life.

Materials and Procedure

• Appartus: 2 porcelain chips, 1 hot plate, 1 funnel holder, 1 water bath
• Chemicals Required for RNA preparation from yeast: 20ml 10% NaOH, 1ml concentrated HCl, 10 g yeast, 35ml 95% ethanol