Molecular Biology Techniques: A Classroom Laboratory Manual, Fourth Edition is a must-have collection of methods and procedures on how to create a single, continuous, comprehensive project that teaches students basic molecular techniques. It is an indispensable tool for introducing advanced undergraduates and beginning graduate students to the techniques of recombinant DNA technology—or gene cloning and expression. The techniques used in basic research and biotechnology laboratories are covered in detail. Students will gain hands-on experience on subcloning a gene into an expression vector straight through to the purification of the recombinant protein.
Key Features
- Presents student-tested labs proven successful in real classroom laboratories
- Includes a test bank on a companion website for additional testing and practice
- Provides exercises that simulate a cloning project that would be performed in a real research lab
- Includes a prep-list appendix that contains necessary recipes and catalog numbers, providing staff with detailed instructions
Part I: Manipulation of DNAAdvanced Alternatives Within Part I 1. Getting Oriented; Practicing With Micropipettes 2. Purification and Digestion of Plasmid (Vector) DNA3. Completion of Vector Preparation and Polymerase Chain Reaction Amplification of egfp 4. Preparation of Insert DNA (egfp) PCR Product5. DNA Ligation and Transformation of Escherichia coli
Part II: Screening TransformantsAdvanced Alternatives Within Part II6. Screening of Transformants, Part I 7. Screening of Transformants, Part II8. Analysis of DNA Sequence From a Positive Clone, Part II
Part III: Expression, Detection, and Purification of Recombinant Proteins from BacteriaAdvanced 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 Column12. Analysis of Purification Fractions
Part IV: Analysis of mRNA Levels13. Total RNA Purification14. Analysis of gst::egfp mRNA Levels by RT-qPCR: Part I15. 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 Expression18. Culturing Mammalian Cells19. Transient Transfection of Mammalian Cells20. RNAi-Mediated Knockdown of EGFP: Part I 21. RNAi-Mediated Knockdown of EGFP: Part II22. RNAi-Mediated Knockdown of EGFP: Part III23. CRISPR-Mediated Knockout of EGFP: Part I24. CRISPR-Mediated Knockout of EGFP: Part II 25. Advanced CRISPR: Part I 26. Advanced CRISPR: Part II27. Interim Laboratory Session 28. Advanced CRISPR: Part IV 29. Advanced CRISPR: Part V30. Advanced CRISPR: Part VI
AppendixA: Equipment B: Prep List C: Preparation of Competent Escherichia coli Cells D: Pre-Lab Questions
- Clark, Molecular Biology, 2e, Academic Cell, Feb 2012, 9780123785947, $135.00
- Farrell, RNA Methodologies, 4e, 2009, 9780123747273, $136.00
- Jankowsky, RNA Helicases (Methods in Enzymology), 2012, 9780123965462, $199.00
Graduate and undergraduate students studying biochemistry, molecular biology, biotechnology, and cell biology
It's great!
Perfektes Buch für die molekulare Biologie.
Una guía muy práctica para tener a mano los principales protocoles de laboratorio en biología molecular. Hay pocas publicaciones técnicas de referencia en este campo y se agradece encontrar una obra de consulta que además sepa explicar el por qué de los distintos métodos.