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Dedication
Acknowledgments
Reviewer's Summary
Introduction
To the Students
To the Instructors
Laboratory Safety
Pipetting
Making Solutions and Dilutions
Weekly Laboratory Notes
Laboratory Report
Overview
Equipment and Materials
Week 1: Genomic DNA Isolation and Quantification; Degenerate PCR
Week 2: Agarose Gel Electrophoresis; Purification and Quantification of PCR Products; Degenerate PCR Primer Design
Week 3: TOPO-based DNA Cloning of Nucleotide Binding Site (NBS) Sequences; Bacterial Transformation
References
Week 1: Plasmid DNA Isolation and Quantification; Restriction Digestion; Sample Preparation for DNA Sequencing
Week 2: Agarose Gel Electrophoresis; Restriction Mapping; Southern Transfer
Week 3: DNA Extraction from Agarose Gel and Quantification; PCR Digoxigenin Probe Synthesis
Week 4: Southern Blot Hybridization and Detection
Week 5: Web-based Sequence Analysis of Cloned NBS Sequences
Week 1: Recombineering of
Week 2: Validation of Recombineering-edited
Week 3: CRISPR-Cas9-assisted gene editing in
Week 4: Validation of CRISPR-Cas9-assisted Gene-editing in
Week 1: Total RNA Isolation, Quantification, and Integrity Assessment
Week 2: RT-qPCR
Week 1: Protein Isolation and Quantification for SDS-polyacrylamide Gel Electrophoresis (SDS-PAGE)
Week 2: SDS-polyacrylamide Gel Electrophoresis (SDS-PAGE) and Western Blotting
Week 3: Immunodetection of Western Blot for Bovine Serum Albumin (BSA) Protein
Appendix A: Working Solutions for Immunodetection of Southern Blot
Appendix B: General Remarks on Handling RNA
Appendix C: Instructor Key for Solutions and Dilutions
About the Author
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Molecular Techniques Copyright © 2026 by Ming-Mei Chang is licensed under a Creative Commons Attribution-ShareAlike 4.0 International License, except where otherwise noted.