Bringing this best-selling textbook right up to date, the new edition uniquely integrates the theories and methods that drive the fields of biology, biotechnology and medicine, comprehensively covering both the techniques students will encounter in lab classes and those that underpin current key advances and discoveries. The contents have been updated to include both traditional and cutting-edge techniques most commonly used in current life science research. Emphasis is placed on understanding the theory behind the techniques, as well as analysis of the resulting data. New chapters cover proteomics, genomics, metabolomics, and bioinformatics, as well as data analysis and visualisation. Using accessible language to describe concepts and methods, and with a wealth of new in-text worked examples to challenge students' understanding, this textbook provides an essential guide to the key techniques used in current bioscience research.
Introduces the latest methods in the life sciences, including cutting-edge techniques such as CRISPR/gene editing and single molecule diffraction/imaging
Promotes problem-solving with lots of worked examples to support students in understanding the quantitative calculations behind the methods
The book is re-organised and extensively rewritten to ensure coverage of all the key current research areas including omics methods, as well as providing the informatics and analytical tools to support students in handling the data generated
Reviews & endorsements
'After seven editions between 1975 and 2010 this essential textbook has undergone a major update. The new editors, Drs Hofmann and Clokie, recruited many experts to revise individual chapters and added a significant amount of new material, thus making the book even more valuable. This eighth edition is basically a new book that covers the techniques of biochemistry and molecular biology in a very comprehensive manner - it does not go into great detail, but gives the reader a good introduction to all of them. Each chapter contains suggestions for further reading for those in need of a follow-up. Excellent diagrams and illustrations provide help in comprehension of the presented material. Since this text is largely aimed at undergraduate students, it fulfils its role extremely well.' Alexander Wlodawer, National Cancer Institute at Frederick
Review from previous edition: ‘This book deserves a place in any modern pathology department and brings together a body of information which is traditionally scattered across several texts … it also represents excellent value for money.’ Annals of Clinical Biochemistry
Review from previous edition: ‘It is necessary to have a practical up to date guide to experimental techniques at hand in a laboratory. This book belongs to such kind of texts. It covers all branches of basic as well as advanced techniques. The book represents a fundamental guidebook for work in a laboratory.’ Photosynthetica
Review from previous edition: ‘The book is well illustrated, indexed, free of errors and most chapters have problem sets with worked answers. The book is written for ‘all bioscience undergraduate students and pre-medical students for whom practical biochemistry, molecular biology and immunology form part of the syllabus’ … it is a very good supplement for undergraduate courses in general biochemistry or a text for courses in biochemical techniques. [This book] is also recommended for graduate students and other researchers in applied sciences such as food biochemistry.’ Journal of Food Biochemistry
Review from previous edition: ‘… well-written and comprehensive … what truly distinguishes this text from others of its kind are the outstanding chapters on proteins, mass spectrometry and spectroscopic techniques. … a valuable resource for undergraduate or graduate students taking introductory or advanced techniques/laboratory courses in cell/molecular biology, biochemistry or biotechnology.‘ Microbiology Today
Review from previous edition: ‘I found this volume remarkably satisfying in that it answered all those 'why do you do it like this?' type questions. … This book has already received extensive reference value on my bookshelf.' Journal of Biological Education
Author(s): Andreas Hofmann, Samuel Clokie
Edition: 8
Publisher: Cambridge University Press
Year: 2018
Language: English
Tags: Molecular Biology; Biochemistry; Structural 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
