Advanced Methods in Molecular Biology and Biotechnology: A Practical Lab Manual is a concise reference on common protocols and techniques for advanced molecular biology and biotechnology experimentation. Each chapter focuses on a different method, providing an overview before delving deeper into the procedure in a step-by-step approach. Techniques covered include genomic DNA extraction using cetyl trimethylammonium bromide (CTAB) and chloroform extraction, chromatographic techniques, ELISA, hybridization, gel electrophoresis, dot blot analysis and methods for studying polymerase chain reactions. Laboratory protocols and standard operating procedures for key equipment are also discussed, providing an instructive overview for lab work.
This practical guide focuses on the latest advances and innovations in methods for molecular biology and biotechnology investigation, helping researchers and practitioners enhance and advance their own methodologies and take their work to the next level.
1. Introduction - p. page_xxvii 2. About the author - p. page_xxix 3. Disclaimer - p. page_xxxi 4. Chapter 1: Introduction to molecular biology techniques - p. page_1 5. Chapter 2: Agarose gel electrophoresis - p. page_7 6. Chapter 3: SDS-polyacrylamide gel electrophoresis - p. page_13 7. Chapter 4: Immunoelectrophoresis - p. page_21 8. Chapter 5: Plasmid DNA isolation using alkaline lysis method from E. coli DH5α cells - p. page_27 9. Chapter 6: Restriction digestion of plasmid DNA - p. page_33 10. Chapter 7: Genomic DNA extraction from the plant leaves using the CTAB method - p. page_37 11. Chapter 8: Genomic DNA extraction from silkworm using modified phenol:chloroform method (Suzuki et al., 1972) - p. page_45 12. Chapter 9: Genomic DNA extraction from Fungal Mycelium using the modified Cenis, 1992 method - p. page_51 13. Chapter 10: Bacterial transformation using CaCl2 - p. page_57 14. Chapter 11: Transformation of E. coli by electroporation - p. page_63 15. Chapter 12: Transfection of cells using lipofectamine - p. page_69 16. Chapter 13: Total RNA isolation - p. page_73 17. Chapter 14: RNA isolation from plant tissues with high phenolic compounds and polysaccharides - p. page_79 18. Chapter 15: Total protein extraction with TCA-acetone and 2D gel electrophoresis - p. page_83 19. Chapter 16: Dansyl-Edman method of peptide sequencing - p. page_89 20. Chapter 17: ISSR analysis - p. page_93 21. Chapter 18: Gene cloning/recombinant DNA technology - p. page_97 22. Chapter 19: Polymerase chain reaction (PCR) - p. page_109 23. Chapter 20: Blotting/hybridization and its types - p. page_117 24. Chapter 21: Dot blot analysis - p. page_123 25. Chapter 22: ELISA (enzyme-linked immunosorbent assay) - p. page_127 26. Chapter 23: Study of principle of centrifugation - p. page_133 27. Chapter 24: Study of principle of chromatography - p. page_139 28. Chapter 25: Thin layer chromatography (TLC) - p. page_143 29. Chapter 26: Gel-filtration or size-exclusion chromatography - p. page_147 30. Chapter 27: Ion-exchange chromatography - p. page_151 31. Chapter 28: To perform affinity chromatography - p. page_155 32. Chapter 29: To perform paper chromatography - p. page_159 33. Chapter 30: Growth of bacterial cultures and preparation of growth curve - p. page_163 34. Chapter 31: To study standard operating procedures (SOPs) for various equipment - p. page_167 35. Index - p. page_181
Advanced Methods in Molecular Biology and Biotechnology: A Practical Lab Manual is a concise reference on common protocols and techniques for advanced molecular biology and biotechnology experimentation. Each chapter focuses on a different method, providing an overview before delving deeper into the procedure in a step-by-step approach. Techniques covered include genomic DNA extraction using cetyl trimethylammonium bromide (CTAB) and chloroform extraction, chromatographic techniques, ELISA, hybridization, gel electrophoresis, dot blot analysis and methods for studying polymerase chain reactions. Laboratory protocols and standard operating procedures for key equipment are also discussed, providing an instructive overview for lab work.
This practical guide focuses on the latest advances and innovations in methods for molecular biology and biotechnology investigation, helping researchers and practitioners enhance and advance their own methodologies and take their work to the next level.
1. Introduction - p. page_xxvii 2. About the author - p. page_xxix 3. Disclaimer - p. page_xxxi 4. Chapter 1: Introduction to molecular biology techniques - p. page_1 5. Chapter 2: Agarose gel electrophoresis - p. page_7 6. Chapter 3: SDS-polyacrylamide gel electrophoresis - p. page_13 7. Chapter 4: Immunoelectrophoresis - p. page_21 8. Chapter 5: Plasmid DNA isolation using alkaline lysis method from E. coli DH5α cells - p. page_27 9. Chapter 6: Restriction digestion of plasmid DNA - p. page_33 10. Chapter 7: Genomic DNA extraction from the plant leaves using the CTAB method - p. page_37 11. Chapter 8: Genomic DNA extraction from silkworm using modified phenol:chloroform method (Suzuki et al., 1972) - p. page_45 12. Chapter 9: Genomic DNA extraction from Fungal Mycelium using the modified Cenis, 1992 method - p. page_51 13. Chapter 10: Bacterial transformation using CaCl2 - p. page_57 14. Chapter 11: Transformation of E. coli by electroporation - p. page_63 15. Chapter 12: Transfection of cells using lipofectamine - p. page_69 16. Chapter 13: Total RNA isolation - p. page_73 17. Chapter 14: RNA isolation from plant tissues with high phenolic compounds and polysaccharides - p. page_79 18. Chapter 15: Total protein extraction with TCA-acetone and 2D gel electrophoresis - p. page_83 19. Chapter 16: Dansyl-Edman method of peptide sequencing - p. page_89 20. Chapter 17: ISSR analysis - p. page_93 21. Chapter 18: Gene cloning/recombinant DNA technology - p. page_97 22. Chapter 19: Polymerase chain reaction (PCR) - p. page_109 23. Chapter 20: Blotting/hybridization and its types - p. page_117 24. Chapter 21: Dot blot analysis - p. page_123 25. Chapter 22: ELISA (enzyme-linked immunosorbent assay) - p. page_127 26. Chapter 23: Study of principle of centrifugation - p. page_133 27. Chapter 24: Study of principle of chromatography - p. page_139 28. Chapter 25: Thin layer chromatography (TLC) - p. page_143 29. Chapter 26: Gel-filtration or size-exclusion chromatography - p. page_147 30. Chapter 27: Ion-exchange chromatography - p. page_151 31. Chapter 28: To perform affinity chromatography - p. page_155 32. Chapter 29: To perform paper chromatography - p. page_159 33. Chapter 30: Growth of bacterial cultures and preparation of growth curve - p. page_163 34. Chapter 31: To study standard operating procedures (SOPs) for various equipment - p. page_167 35. Index - p. page_181
