Wilson and Walker's Principles and Techniques of Biochemistry and Molecular Biology

Cover
Title Page
Copyright
Contents
Foreword to the Eighth Edition
Preface
List of Contributors
Tables and Resources
1 Biochemical and Molecular Biological Methods in Life Sciences Studies
	1.1 From Biochemistry and Molecular Biology to the Life Sciences
	1.2 The Education of Life Scientists
	1.3 Aims of Life Science Studies
	1.4 Personal Qualities and Scientific Conduct
	1.5 Suggestions for Further Reading
2 Basic Principles
	2.1 Biologically Important Molecules
	2.2 The Importance of Structure
	2.3 Parameters of Biological Samples
	2.4 Measurement of the pH: The pH Electrode
	2.5 Buffers
	2.6 Ionisation Properties of Amino Acids
	2.7 Quantitative Biochemical Measurements
	2.8 Experiment Design and Research Conduct
	2.9 Suggestions for Further Reading
3 Cell Culture Techniques
	3.1 Introduction
	3.2 The Cell Culture Laboratory and Equipment
	3.3 Safety Considerations in Cell Culture
	3.4 Aseptic Techniques and Good Cell Culture Practice
	3.5 Types of Animal Cells, Characteristics and Maintenance in Culture
	3.6 Stem Cell Culture
	3.7 Bacterial Cell Culture
	3.8 Potential Use of Cell Cultures
	3.9 Acknowledgements
	3.10 Suggestions for Further Reading
4 Recombinant DNA Techniques and Molecular Cloning
	4.1 Introduction
	4.2 Structure of Nucleic Acids
	4.3 Genes and Genome Complexity
	4.4 Location and Packaging of Nucleic Acids
	4.5 Functions of Nucleic Acids
	4.6 The Manipulation of Nucleic Acids: Basic Tools and Techniques
	4.7 Isolation and Separation of Nucleic Acids
	4.8 Automated Analysis of Nucleic Acid Fragments
	4.9 Molecular Analysis of Nucleic Acid Sequences
	4.10 The Polymerase Chain Reaction (PCR)
	4.11 Constructing Gene Libraries
	4.12 Cloning Vectors
	4.13 Hybridisation and Gene Probes
	4.14 Screening Gene Libraries
	4.15 Applications of Gene Cloning
	4.16 Expression of Foreign Genes
	4.17 Analysing Genes and Gene Expression
	4.18 Analysing Genetic Mutations and Polymorphisms
	4.19 Molecular Biotechnology and Applications
	4.20 Pharmacogenomics
	4.21 Suggestions for Further Reading
5 Preparative Protein Biochemistry
	5.1 Introduction
	5.2 Determination of Protein Concentrations
	5.3 Engineering Proteins for Purification
	5.4 Producing Recombinant Protein
	5.5 Cell-Disruption Methods
	5.6 Preliminary Purification Steps
	5.7 Principles of Liquid Chromatography
	5.8 Chromatographic Methods for Protein Purification
	5.9 Other Methods of Protein Purification
	5.10 Monitoring Protein Purification
	5.11 Storage
	5.12 Suggestions for Further Reading
6 Electrophoretic Techniques
	6.1 General Principles
	6.2 Support Media and Buffers
	6.3 Electrophoresis of Proteins
	6.4 Electrophoresis of Nucleic Acids
	6.5 Capillary Electrophoresis
	6.6 Microchip Electrophoresis
	6.7 Suggestions for Further Reading
7 Immunochemical Techniques
	7.1 Introduction
	7.2 Antibody Preparation
	7.3 Immunoassay Formats
	7.4 Immuno Microscopy
	7.5 Lateral Flow Devices
	7.6 Epitope Mapping
	7.7 Immunoblotting
	7.8 Fluorescence-Activated Cell Sorting (FACS)
	7.9 Cell and Tissue Staining Techniques
	7.10 Immunocapture Polymerase Chain Reaction (PCR)
	7.11 Immunoaffinity Chromatography
	7.12 Antibody-Based Biosensors
	7.13 Luminex® Technology
	7.14 Therapeutic Antibodies
	7.15 Suggestions for Further Reading
8 Flow Cytometry
	8.1 Introduction
	8.2 Instrumentation
	8.3 Fluorescence-Activated Cell Sorting (FACS)
	8.4 Fluorescence Labels
	8.5 Practical Considerations
	8.6 Applications
	8.7 Suggestions for Further Reading
9 Radioisotope Techniques
	9.1 Why Use a Radioisotope?
	9.2 The Nature of Radioactivity
	9.3 Detection and Measurement of Radioactivity
	9.4 Other Practical Aspects of Counting Radioactivity and Analysis of Data
	9.5 Safety Aspects
	9.6 Suggestions for Further Reading
10 Principles of Clinical Biochemistry
	10.1 Principles of Clinical Biochemical Analysis
	10.2 Clinical Measurements and Quality Control
	10.3 Examples of Biochemical Aids to Clinical Diagnosis
	10.4 Suggestions for Further Reading
11 Microscopy
	11.1 Introduction
	11.2 The Light Microscope
	11.3 Optical Sectioning
	11.4 Imaging Live Cells and Tissues
	11.5 Measuring Cellular Dynamics
	11.6 The Electron Microscope
	11.7 Image Management
	11.8 Suggestions for Further Reading
12 Centrifugation and Ultracentrifugation
	12.1 Introduction
	12.2 Basic Principles of Sedimentation
	12.3 Types, Care and Safety Aspects of Centrifuges
	12.4 Preparative Centrifugation
	12.5 Analytical Ultracentrifugation
	12.6 Suggestions for Further Reading
13 Spectroscopic Techniques
	13.1 Introduction
	13.2 Ultraviolet and Visible Light Spectroscopy
	13.3 Circular Dichroism Spectroscopy
	13.4 Infrared and Raman Spectroscopy
	13.5 Fluorescence Spectroscopy
	13.6 Luminometry
	13.7 Atomic Spectroscopy
	13.8 Rapid Mixing Techniques for Kinetics
	13.9 Suggestions for Further Reading
14 Basic Techniques Probing Molecular Structure and Interactions
	14.1 Introduction
	14.2 Isothermal Titration Calorimetry
	14.3 Techniques to Investigate the Three-Dimensional Structure
	14.4 Switch Techniques
	14.5 Solid-Phase Binding Techniques with Washing Steps
	14.6 Solid-Phase Binding Techniques Combined with Flow
	14.7 Suggestions for Further Reading
15 Mass Spectrometric Techniques
	15.1 Introduction
	15.2 Ionisation
	15.3 Mass Analysers
	15.4 Detectors
	15.5 Other Components
	15.6 Suggestions for Further Reading
16 Fundamentals of Bioinformatics
	16.1 Introduction
	16.2 Biological Databases
	16.3 Biological Data Formats
	16.4 Sequence Alignment and Tools
	16.5 Annotation of Predicted Peptides
	16.6 Principles of Phylogenetics
	16.7 Suggestions for Further Reading
17 Fundamentals of Chemoinformatics
	17.1 Introduction
	17.2 Computer Representations of Chemical Structure
	17.3 Calculation of Compound Properties
	17.4 Molecular Mechanics
	17.5 Databases
	17.6 Suggestions for Further Reading
18 The Python Programming Language
	18.1 Introduction
	18.2 Getting Started
	18.3 Examples
	18.4 Suggestions for Further Reading
19 Data Analysis
	19.1 Introduction
	19.2 Data Representations
	19.3 Data Analysis
	19.4 Conclusion
	19.5 Suggestions for Further Reading
20 Fundamentals of Genome Sequencing and Annotation
	20.1 Introduction
	20.2 Genomic Sequencing
	20.3 Assembly of Genomic Information
	20.4 Prediction of Genes
	20.5 Functional Annotation
	20.6 Post-Genomic Analyses
	20.7 Factors Affecting the Sequencing, Annotation and Assembly of Eukaryotic Genomes, and Subsequent Analyses
	20.8 Concluding Remarks
	20.9 Suggestions For Further Reading
21 Fundamentals of Proteomics
	21.1 Introduction: From Edman Sequencing to Mass Spectrometry
	21.2 Digestion
	21.3 Tandem Mass Spectrometry
	21.4 The Importance of Isotopes for Finding the Charge State of a Peptide
	21.5 Sample Preparation and Handling
	21.6 Post-Translational Modification of Proteins
	21.7 Analysing Protein Complexes
	21.8 Computing and Database Analysis
	21.9 Suggestions for Further Reading
22 Fundamentals of Metabolomics
	James I. MacRae 22.1 Introduction: What is Metabolomics?
	22.2 Sample Preparation
	22.3 Data Acquisition
	22.4 Untargeted and Targeted Metabolomics
	22.5 Chemometrics and Data Analysis
	22.6 Further Metabolomics Techniques and Terminology
	22.7 Suggestions for Further Reading
23 Enzymes and Receptors
	23.1 Definition and Classification of Enzymes
	23.2 Enzyme Kinetics
	23.3 Analytical Methods to Investigate Enzyme Kinetics
	23.4 Molecular Mechanisms of Enzymes
	23.5 Regulation of Enzyme Activity
	23.6 Receptors
	23.7 Characterisation of Receptor–Ligand Binding
	23.8 Suggestions for Further Reading
24 Drug Discovery and Development
	24.1 Introduction
	24.2 Molecular Libraries and Drug-Discovery Strategies
	24.3 Assembling a Molecular Library
	24.4 Compound Management
	24.5 Screening Strategies Used in Hit Discovery
	24.6 Active-to-Hit Phase
	24.7 Hit-to-Lead Phase
	24.8 ADMET
	24.9 Lead Optimisation
	24.10 Suggestions for Further Reading
Index