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| Theory and Experimental Techniques | |
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| Introduction to the Biochemistry Laboratory | |
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| Safety in the Laboratory | |
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| The Laboratory Notebook and Experiment Reports | |
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| Details of Experimental Write-up | |
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| Cleaning Laboratory Glassware | |
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| Glassware | |
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| Quartz and Glass Cuvettes | |
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| Preparation and Storage of Solutions | |
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| Water Quality | |
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| Solution Preparation | |
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| Quantitative Transfer of Liquids | |
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| Filling a Pipet | |
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| Disposable Pasteur Pipets | |
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| Calibrated Pipets | |
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| Automatic Pipetting Systems | |
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| Cleaning and Drying Pipets | |
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| Statistical Analysis of Experimental Data | |
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| Analysis of Experimental Data | |
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| Determination of the Mean, Sample Deviation, and Standard Deviation | |
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| Statistical Analysis in Practice | |
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| Study Problems | |
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| Further Reading | |
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| On the Web | |
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| General Laboratory Procedures | |
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| pH, Buffers, Electrodes, and Biosensors | |
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| Measurement of pH | |
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| Biochemical Buffers | |
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| Selection of a Biochemical Buffer | |
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| The Oxygen Electrode | |
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| Ion-Selective Electrodes and Biosensors | |
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| Measurement of Protein Solutions | |
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| The Biuret and Lowry Assays | |
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| The Bradford Assay | |
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| The BCA Assay | |
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| The Spectrophotometric Assay | |
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| Measurement of Nucleic Acid Solutions | |
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| The Spectrophotometric Assay | |
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| Other Assays for Nucleic Acids | |
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| Techniques for Sample Preparation | |
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| Dialysis | |
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| Ultrafiltration | |
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| Lyophilization and Centrifugal Vacuum Concentration | |
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| Study Problems | |
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| Further Reading | |
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| On the Web | |
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| Purification and Identification of Biomolecules by Chromatography | |
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| Introduction to Chromatography | |
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| Partition versus Adsorption Chromatography | |
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| Planar Chromatography (Paper and Thin-Layer) | |
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| Preparation of the Stationary Support | |
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| Solvent Development of the Support | |
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| Detection and Measurement of Components | |
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| Applications of Planar Chromatography | |
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| Gas Chromatography (GC) | |
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| Instrumentation | |
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| Selection of Operating Conditions | |
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| Analysis of GC Data | |
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| Advantages and Limitations of GC | |
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| Column Chromatography | |
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| Operation of a Chromatographic Column | |
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| Packing the Column | |
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| Loading the Column | |
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| Eluting the Column | |
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| Collecting the Eluent | |
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| Detection of Eluting Components | |
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| Ion-Exchange Chromatography | |
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| Ion-Exchange Resins | |
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| Selection of the Ion Exchanger | |
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| Choice of Buffer | |
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| Preparation of the Ion Exchanger | |
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| Using the Ion-Exchange Resin | |
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| Storage of Resins | |
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| Gel Exclusion Chromatography | |
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| Theory of Gel Filtration | |
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| Physical Characterization of Gel Chromatography | |
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| Chemical Properties of Gels | |
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| Selecting a Gel | |
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| Gel Preparation and Storage | |
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| Operation of a Gel Column | |
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| Applications of Gel Exclusion Chromatography | |
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| High-Performance Liquid Chromatography (HPLC) | |
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| Instrumentation | |
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| Stationary Phases in HPLC | |
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| The Mobile Phase | |
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| Solvents for HPLC Operation | |
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| Gradient Elution in HPLC | |
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| Sample Preparation and Selection of HPLC Operating Conditions | |
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| FPLC--A Modification of HPLC | |
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| Affinity Chromatography and Immunoadsorption | |
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| Chromatographic Media | |
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| The Immobilized Ligand | |
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| Attachment of Ligand to Matrix | |
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| Experimental Procedure for Affinity Chromatography | |
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| Perfusion Chromatography | |
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| Study Problems | |
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| Further Reading | |
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| Chromatography on the Web | |
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| Characterization of Proteins and Nucleic Acids by Electrophoresis | |
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| Theory of Electrophoresis | |
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| Methods of Electrophoresis | |
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| Polyacrylamide Gel Electrophoresis (PAGE) | |
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| Nucleic Acid Sequencing Gels | |
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| Agarose Gel Electrophoresis | |
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| Pulsed Field Gel Electrophoresis (PFGE) | |
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| Isoelectric Focusing of Proteins | |
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| Two-Dimensional Electrophoresis of Proteins | |
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| Capillary Electrophoresis (CE) | |
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| Immunoelectrophoresis (IE) | |
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| Practical Aspects of Electrophoresis | |
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| Instrumentation | |
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| Reagents | |
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| Staining and Detecting Electrophoresis Bands | |
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| Protein and Nucleic Acid Blotting | |
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| Analysis of Electrophoresis Results | |
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| Study Problems | |
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| Further Reading | |
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| Electrophoresis on the Web | |
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| Spectroscopic Analysis of Biomolecules | |
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| Ultraviolet-Visible Absorption Spectrophotometry | |
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| Principles | |
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| Instrumentation | |
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| Applications | |
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| Fluorescence Spectrophotometry | |
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| Principles | |
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| Instrumentation | |
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| Applications | |
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| Difficulties in Fluorescence Measurements | |
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| Other Spectroscopic Methods | |
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| Nuclear Magnetic Resonance Spectroscopy | |
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| Mass Spectrometry | |
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| Study Problems | |
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| Further Reading | |
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| Spectroscopy on the Web | |
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| Radioisotopes in Biochemical Research | |
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| Origin and Properties of Radioactivity | |
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| Introduction | |
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| Isotopes in Biochemistry | |
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| Units of Radioactivity | |
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| Detection and Measurement of Radioactivity | |
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| Liquid Scintillation Counting | |
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| Geiger-Muller Counting of Radioactivity | |
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| Scintillation Counting of [gamma] Rays | |
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| Background Radiation | |
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| Applications of Radioisotopes | |
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| Statistical Analysis of Radioactivity Measurements | |
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| Radioisotopes and Safety | |
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| Preparation for the Experiment | |
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| Performing the Experiment | |
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| Study Problems | |
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| Further Reading | |
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| Radioisotopes on the Web | |
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| Centrifugation in Biochemical Research | |
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| Basic Principles of Centrifugation | |
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| Instrumentation for Centrifugation | |
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| Low-Speed Centrifuges | |
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| High-Speed Centrifuges | |
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| Ultracentrifuges | |
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| Applications of Centrifugation | |
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| Preparative Techniques | |
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| Analytical Measurements | |
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| Care of Centrifuges and Rotors | |
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| Study Problems | |
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| Further Reading | |
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| Centrifugation on the Web | |
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| Experiments | |
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| Using the Computer in Biochemical Research | |
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| Structural Analysis of a Dipeptide | |
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| Using Gel Filtration to Study Ligand-Protein Interactions | |
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| Isolation and Characterization of Bovine Milk [alpha]-Lactalbumin | |
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| Kinetic Analysis of Tyrosinase | |
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| Purification and Characterization of Triacylglycerols in Natural Oils | |
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| Identification of Serum Glycoproteins by SDS-PAGE and Western Blotting | |
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| Isolation and Characterization of Plant Pigments | |
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| Photoinduced Proton Transport through Chloroplast Membranes | |
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| Isolation, Subfractionation, and Enzymatic Analysis of Beef Heart Mitochondria | |
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| Measurement of Cholesterol and Vitamin C in Biological Samples | |
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| Activity and Thermal Stability of Gel-immobilized Peroxidase | |
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| Extraction and Characterization of Bacterial DNA | |
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| Plasmid DNA Isolation and Characterization by Electrophoresis | |
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| The Action of Restriction Endonucleases on Plasmid or Viral DNA | |
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| Appendices | |
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| Properties of Common Acids and Bases | |
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| Properties of Common Buffer Compounds | |
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| pK[subscript a] Values and pH[subscript I] Values of Amino Acids | |
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| Molecular Weights of Some Common Proteins | |
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| Common Abbreviations Used in This Text | |
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| Units of Measurement | |
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| Table of the Elements | |
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| Values of t for Analysis of Statistical Confidence Limits | |
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| Answers to Selected Study Problems | |
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| Index | |