Molecular Biology Techniques : A Classroom Laboratory Manual

Part I: Manipulation of DNA Advanced Alternatives Within Part I 1. Getting Oriented; Practicing With Micropipettes 2. Purification and Digestion of Plasmid (Vector) DNA 3. Completion of Vector Preparation and Polymerase Chain Reaction Amplification of egfp 4. Preparation of Insert DNA (egfp) PCR Product 5. DNA Ligation and Transformation of Escherichia coli Part II: Screening Transformants Advanced Alternatives Within Part II 6. Screening of Transformants, Part I 7. Screening of Transformants, Part II 8. Analysis of DNA Sequence From a Positive Clone, Part II Part III: Expression, Detection, and Purification of Recombinant Proteins from Bacteria Advanced Alternatives Within Part III 9. Expression of Fusion Protein from Positive Clones, SDS_PAGE and Western Blot: Part I 10. Expression of Fusion Protein from Positive Clones, SDS_PAGE, and Western Blot: Part II 11. Extraction of Recombinant Protein From Escherichia coli Using a Glutathione Affinity Column 12. Analysis of Purification Fractions Part IV: Analysis of mRNA Levels 13. Total RNA Purification 14. Analysis of gst::egfp mRNA Levels by RT-qPCR: Part I 15. Analysis of gst::egfp mRNA Levels by RT-qPCR: Part II 16. Analysis of gst::egfp mRNA Levels by Semiquantitative RT-PCR: Part I 17. Analysis of gst::egfp mRNA Levels by Semiquantitative RT-PCR: Part II Part V: Modulation of Gene Expression 18. Culturing Mammalian Cells 19. Transient Transfection of Mammalian Cells 20. RNAi-Mediated Knockdown of EGFP: Part I 21. RNAi-Mediated Knockdown of EGFP: Part II 22. RNAi-Mediated Knockdown of EGFP: Part III 23. CRISPR-Mediated Knockout of EGFP: Part I 24. CRISPR-Mediated Knockout of EGFP: Part II 25. Advanced CRISPR: Part I 26. Advanced CRISPR: Part II 27. Interim Laboratory Session 28. Advanced CRISPR: Part IV 29. Advanced CRISPR: Part V 30. Advanced CRISPR: Part VI Appendix A: Equipment B: Prep List C: Preparation of Competent Escherichia coli Cells D: Pre-Lab Questions

9780128180242

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