Molecular Biology and Biotechnology: basic experimental protocols is a compilation of methods and techniques commonly used in biomedical and biotechnological studies. The book aims to provide ample support to both students and faculty while conducting practical lessons. Four sections are covered in this book—Genomics, Proteomics, Quantitative Biochemistry, and Bioinformatics. A concise introductory note accompanies each protocol/method described for better comprehension. Every topic discussed is supported by actual methods and their expected results, and is accompanied by relevant questions.

Table of Contents:
SECTION I: GENOMICS 1. Initial Characterization of DNA 1.1 Mammalian High Molecular Weight DNA Extraction 1.2 Spectrophotometric Analysis of DNA 1.3 Agarose Gel Electrophoretic Analysis of DNA 1.4 Restriction Mapping of DNA 2. DNA Probe Preparation, Southern Blotting, and Hybridization 2.1 Oligonucleotides or Gene Probe Design 2.2 Labelling DNA Gene Probes 2.3 Southern Blotting of DNA and Hybridization 3. RFLP and SSCP Analysis of DNA 3.1 Restriction Fragment Length Polymorphism 3.2 Single-strand Conformation Polymorphism 4. Protein–DNA Interaction 4.1 DNase Footprinting of DNA 4.2 Gel Mobility Shift Assay 5. Plasmid DNA Analysis 5.1 Preparation and Maintenance of Microbial Culture 5.2 Plasmid and Chromosomal DNA Preparation from E. coli 5.3 Analysis of Plasmid and Chromosomal DNA on AgaroseGel Electrophoresis 5.4 Transformation of E. coli with Plasmid DNA 5.5 DNA Ligation and E. coli Transformation 6. Polymerase Chain Reaction 6.1 Amplification of DNA by PCR and Analysis on AgaroseGel Electrophoresis 6.2 Cloning PCR Products 6.3 DNA Sequencing 7. RNA Analysis Techniques 7.1 Total RNA Isolation from Tissue or Cells 7.2 Spectrophotometric Analysis of RNA 7.3 RNA Agarose Gel Electrophoresis 7.4 Messenger RNA Extraction from Total RNA 7.5 In Vitro Protein Synthesis 8. Gene or mRNA Expression in Cells/Tissues 8.1 Northern Blotting of RNA and Hybridization 8.2 Reverse Transcriptase-Polymerase Chain Reaction 8.3 Differential Display Polymerase Chain Reaction 8.4 In Situ Hybridization 9. Transcription Activity In Vivo 9.1 Nuclear Run-on Transcription 9.2 Reporter Gene Transcription 10. Apoptosis Analysis 10.1 DNA Ladder 10.2 TUNEL Assay 10.3 Comet Assay SECTION II: PROTEOMICS 11. Initial Preparations for Protein Analysis 11.1 Buffers and Additional Components 11.2 Spectroscopic Analysis of Chromophores 11.3 Tissue and their Subcellular Fractionation 12. Quantitative Determination of Proteins 12.1 Lowry (Folin–Ciocalteu) Method 12.2 Bradford Method 12.3 Direct Spectrophotometric Method 13. Protein Separation and Analysis 13.1 Ion-exchange Chromatography 13.2 Gel Filtration Chromatography 13.3 Electrophoretic Analysis 13.4 Staining of Gels after Electrophoresis 14. Electroblotting, Dot/Slot Blotting, and Electroelution 14.1 Electroblotting 14.2 Dot/Slot Blotting 14.3 Staining Methods of Proteins on Electroblots 14.4 Electroelution of Proteins from Gels 14.5 Sample Preparation for Amino Acid Sequencing 15. Polyclonal Antibodies Production and Immunoanalysis 15.1 Production of Antibodies 15.2 Ouchterlony Double Immunodiffusion Method 15.3 Western Immunoblot 15.4 Enzyme-linked Immunosorbent Assay 15.5 Dot/Slot Immunoblot Assay 15.6 Immunoprecipitation 15.7 Immunohistochemistry 16. Protein–protein Interactions 16.1 Pull-Down Assay 16.2 Far-western Analysis 16.3 Co-immunoprecipitation SECTION III: QUANTITATIVE BIOCHEMISTRY 17. Analysis of Biomolecules 17.1 Extraction and Estimation of DNA, RNA, and Protein from Tissues 17.2 Estimation of Oxidative Stress-related Biomolecules 18. Enzyme Kinetics and Enzyme Activity Analysis 18.1 Influence of Substrate Concentration on Enzyme Activity 18.2 Influence of Temperature on Enzyme Activity 18.3 Influence of pH on Enzyme Activity 18.4 Influence of Inhibitors on Enzyme Activity 18.5 Redox-active Enzyme Activity Estimations 19. Labelling of Proteins and its Uses 19.1 Radioiodination of Target Protein 19.2 In Vivo Receptor Assay Using 125I-labelled Protein 19.3 Radioactive Phosphorylation of Specific Protein (Tissue Protein Kinase Assay) 20. Statistical Analysis of Biochemical Data 20.1 t-Test 20.2 Analysis of Variance SECTION IV: BIOINFORMATICS 21. Sequence Retrieval from Databases 21.1 Sequence Retrieval 21.2 Using Entrez Search Tags and Boolean Operators 21.3 FASTA Format 22. Sequ