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Molecular biology and biotechnology: basic experimental protocols

  • Author(s): M P Bansal
  • Pages: 392
  • Binding: Paperback
  • Size: 180mm x 240mm
  • Language: English
  • ISBN: 9788179933794
  • Cover Price:

    Rs.695.00 / US $42.00

  • Special Price: Rs. 625.00 / US $38.00

Book Details

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.

Key Features

  • Ready design of basic experiments in biomedical and biotechnology sciences.
  • Extensive coverage of basic equipment used in biomedical and biotechnological studies.
  • Concise information on each subject, with focus on practical lessons.

Target Audience

    Graduate students, Postgraduate students, Technicians of Biomedical Science, Biotechnology

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. Sequence Similarity Search
    22.1 Homologous Sequences
    22.2 Pairwise Sequence Alignment
    22.3 Multiple Sequence Alignment

    23. Protein Sequence and Structure Analysis
    23.1 Primary Protein Structure Analysis
    23.2 Secondary Structure Prediction and Analysis

    24. Nucleotide Sequence Analysis
    24.1 Gene Prediction
    24.2 Splice Site Prediction
    24.3 Restriction Mapping of DNA
    24.4 PCR Primer Designing

    25. Molecular Phylogeny
    25.1 Preparing Alignment File
    25.2 Distance and Character-based Analysis
    25.3 Using Tree Construction Matrices
    25.4 Plotting Phylogenetic Tree

Keywords

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