Biochemistry laboratory : modern theory and techniques / Rodney Boyer. — 2nd ed. — Upper Saddle River, N.J. : Pearson Prentice Hall, c2012. – (58.173057/B791/2nd ed.)

Contents

    CONTENTS
    
    Preface xiii
    Acknowledgments xvii
    About the Author xix
    Chapter 1 INTRODUCTION TO THE BIOCHEMISTRY LABORATORY 1
    A. Safety in the Laboratory 2
    B. Keeping Records and Communicating Experimental Results 6
    C. Using Biochemical Reagents and Solutions 14
    D. Quantitative Transfer of Liquids 18
    E. Statistical Analysis of Experimental Data 23
    Chapter 2 USING THE COMPUTER AND INTERNET FOR RESEARCH IN BIOCHEMISTRY 35
    A. What Is Research and How Is It Done in Biochemistry? 35
    B. Using Computers in Biochemistry 38
    C. Web Sites Useful in Biochemistry 40
    Chapter 3 GENERAL LABORATORY PROCEDURES 53
    A. pH， Buffers， Electrodes， and Biosensors 53
    B. Measurement of Protein Solutions 67
    C. Measurement of Nucleic Acid Solutions 71
    D. Techniques for Sample Preparation 73
    E. Radioisotopes in Biochemistry 80
    Chapter 4 CENTRIFUGATION TECHNIQUES IN BIOCHEMISTRY 95
    A. Basic Principles of Centrifugation 96
    B. Instrumentation for Centrifugation 99
    C. Applications for Centrifugation 106
    Chapter 5 PURIFICATION AND ANALYSIS OF BIOMOLECULES BY CHROMATOGRAPHY 115
    A. Introduction to Chromatography 116
    B. Planar Chromatography (Paper and Thin-Layer Chromatography) 118
    C. Column Chromatography 122
    D. Ion-Exchange Chromatography 126
    E. Gel-Exclusion Chromatography 132
    F. High-Performance Liquid Chromatography (HPLC) 140
    G. Affinity Chromatography and Immunoadsorption 152
    H. Membrane-Based Chromatography 159
    Chapter 6 CHARACTERIZATION OF PROTEINS AND NUCLEIC ACIDS BY ELECTROPHORESlS 165
    A. The Theory of Electrophoresis 166
    B. Methods of Electrophoresis 167
    C. Practical Aspects of Electrophoresis 188
    Chapter 7 SPECTROSCOPIC ANALYSIS OF BIOMOLECULES 201
    A. Ultraviolet-Visible Absorption Spectrometry 202
    B. Fluorescence Spectrometry 220
    C. Nuclear Magnetic Resonance Spectroscopy 225
    D. Mass Spectrometry 230
    E. X-Ray Crystallography 233
    Chapter 8 BIOMOLECULAR INTERACTIONS: LIGAND BINDING AND ENZYME REACTIONS 239
    A. Ligand-Macromolecule Interactions (Molecular Recognition) 239
    B. Biological Catalysis (Enzymes) 250
    Chapter 9 MOLECULAR BIOLOGY h STRUCTURES AND ANALYSIS OF NUCLEIC ACIDS 267
    A. Introduction to the Nucleic Acids 268
    B. Laboratory Methods for Investigation of DNA and RNA 275
    Chapter 10 MOLECULAR BIOLOGY I1: RECOMBINANT DNA， MOLECULAR CLONING， AND ENZYMOLOGY 289
    A. Recombinant DNA Biotechnology 290
    B. Important Enzymes in Molecular Biology and Biotechnology 297
    C. Nucleic Acid Blotting 304
    Chapter 11 PROTEIN PRODUCTION， PURIFICATION， AND CHARACTERIZATION 307
    A. Procedures for the Purification of Proteins 308
    B. Production of Proteins by Expression of Foreign Genes 317
    C. Protein Characterization 322
    D. Determination of Primary Structure 323
    Appendix I List of Software Programs and Web Sites Useful for Each Chapter 331
    Appendix II Properties of Common Acids and Bases 334
    Appendix III Properties of Common Buffer Compounds 335
    Appendix IV PKa Values and PHI Values of Amino Acids 337
    Appendix V Molecular Weight of Some Common Proteins 338
    Appendix VI Common Abbreviations Used in This Text 339
    Appendix VII Units of Measurement 342
    Appendix VIII Table of the Elements 344
    Appendix IX Answers to Odd-Numbered Study Problems 348
    Index 353