Aim

The determination of the quality of nucleic acid (free from protein contamination or RNA/DNA contamination) and accurate concentration of nucleic acid is important for downstream applications. Techniques like transfection, cloning, polymerase chain reaction (PCR), and next-generation sequencing (NGS), all require high-quality nucleic acid. These applications often have a specific target; a nanogram range of nucleic acid concentration for optimal performance. Inaccurate quantification can increase variability in downstream assays and affect the quality of results.

After completing this experiment, the student will be able to:

1. Understand (Comprehension): Explain the principles of UV absorbance and gel electrophoresis used in nucleic acid quantification.
2. Apply (Application): Use absorbance readings to determine DNA/RNA concentration and purity.
3. Analyze (Analysis): Evaluate nucleic acid integrity by interpreting gel electrophoresis band patterns.
4. Evaluate (Evaluation): Assess contamination in samples based on A₂₆₀/A₂₈₀ and A₂₆₀/A₂₃₀ ratios.
5. Create (Synthesis): Design an experimental workflow combining UV absorbance and gel electrophoresis for quality control of nucleic acids.