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.
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Table of Contents
1. Introduction to molecular biology techniques2. Agarose gel electrophoresis3. SDS-polyacrylamide gel electrophoresis4. Immunoelectrophoresis5. DNA plasmid isolation using Alkaline Lysis Method from E. coli DH5a cells6. Restriction digestion of Plasmid DNA7. Genomic DNA extraction from plant leaves using CTAB method8. Genomic DNA extraction from silkworm using modified phenol: chloroform method (Suzuki et al., 1972)9. Genomic DNA extraction from Fungal Mycelium using modified Cenis, 1992 method10. Bacterial transformation protocol using CaCl2
11. Transformation of E. coli by electroporation12. Transfection of cells using lipofectamine13. Total RNA isolation14. RNA isolation from plant tissues with high phenolic compounds and polysaccharides15. Total protein extraction with TCA-acetone and 2D gel electrophoresis16. Dansyl-Edman method of peptide sequencing17. ISSR analysis18. Gene cloning/recombinant DNA technology19. Polymerase Chain Reaction (PCR)20. Blotting/ hybridization and its types21. Dot blot analysis22. ELISA (enzyme-linked immunosorbent assay)23. Study of principle of centrifugation
24. Study of principle of chromatography
25. Thin layer chromatography (TLC)
26. Gel-filtration or size-exclusion chromatography
27. Ion-exchange chromatography
28. To perform affinity chromatography
29. To perform paper chromatography
30. Growth of bacterial cultures and preparation of growth curve
31. To study standard operating procedures (SOPs) for various equipment