Molecular Techniques: A Project-Based Undergraduate Laboratory

• Introduction
• To the Students
• To the Instructors
• Laboratory Safety
• Pipetting
• Making Solutions and Dilutions
• Weekly Laboratory Notes
• Laboratory Report
• Module 1: Degenerate PCR and Topoisomerase (TOPO)-based DNA Cloning
• Module 2: Analyzing Cloned NBS Sequences
• Module 3: Gene Editing by Recombineering With or Without CRISPR-Cas9
• Module 4: Studying Gene Expressions by Reverse Transcription Quantitative Polymerase Chain Reaction (RT-qPCR)
• Module 5: Exploring the Sensitivity and Specificity of Western Blot
• Appendix A: Working Solutions for Immunodetection of Southern blot
• Appendix B: General Remarks on Handling RNA
• Appendix C: Instructor Key for Solutions and Dilutions

Molecular biology, examining the structures and functions of nucleic acids and proteins, is crucial to advancing biological science and improving human life, particularly in agriculture and medicine. Advances in the field can transform a crop to resist disease or uncover clues to fight illness by manipulating DNA. Techniques for molecular biology research and applications have advanced rapidly over the past decades, and undergraduates need hands-on experiences in this vital field of science. This text provides a structured approach to learning and practicing those techniques, covering DNA, RNA, and protein across five modules and is tailored for a one-semester undergraduate lab course. Each module extends 2–5 weeks of lab exercises and features.