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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 | |