Principles and Techniques of Practical Biochemistry

Keith Wilson, John Walker

Cambridge University Press, Mar 16, 2000 - Medical - 784 pages

In this new, 5th edition of a highly popular text, undergraduate students are introduced to all the basic experimental techniques routinely used in practical biochemistry today. Most attention is given to techniques students will encounter in their practical classes, with the principles and theories behind them explained in detail to aid understanding. As a further aid to students, essential calculations and worked answers appear at the end of each chapter. 'Key terms to understand' are also included to help students thoroughly review each topic. No contemporary book on modern biochemical techniques would be complete without chapters on molecular biology, recombinant DNA technology, genetic analysis and biomolecular interactions, and these topics have been extensively covered in this new edition. The book is essential reading for all bioscience undergraduate students and pre-clinical medical students for whom practical biochemistry, molecular biology and immunology form part of the syllabus.

Contents

• GENERAL PRINCIPLE OF BIOCHEMICAL INVESTIGATIONS: 1
• 12 BIOENERGETICS: 4
• 13 METHODS FOR INVESTIGATING METABOLISM: 21
• 14 PRACTICAL CONSIDERATIONS: 28
• 15 IN VIVO MODUS: 42
• 16 IN VITRO MODELS: 44
• 17 MICROSCOPY: 66
• 18 KEY TERMS: 75
• 19 CALCULATIONS: 76
• 110 SUGGESTIONS FOR FURTHER READING: 78
• MOLECULAR BIOLOGY AND BASIC TECHNIQUES: 80
• 23 GENES AND GENOME COMPLEXITY: 87
• 24 THE NATURE OF THE GENETIC CODE: 90
• 26 THE CELLULAR FUNCTIONS OF DNA: 93
• BASIC TOOLS AND TECHNIQUES: 103
• 28 ISOLATION AND SEPARATION OF NUCLEIC ACIDS: 105
• 29 RESTRICTION MAPPING OF DNA FRAGMENTS: 110
• 210 NUCLEIC ACID BLOTTING METHODS: 111
• 211 GENE PROBE DERIVATION: 113
• 212 LABELLING DNA GENE PROBE MOLECULES: 114
• 213 THE POLYMERASE CHAIN REACTION: 116
• 214 NUCLEOTIDE SEQUENCING OF DNA: 125
• 215 BIOINFORMATICS AND THE INTERNET: 131
• 216 KEY TERMS: 134
• 217 CALCULATIONS: 135
• 218 SUGGESTIONS FOR FURTHER READING: 136
• MOLECULAR CLONING AND GENE ANALYSIS: 138
• 33 CLONING VECTORS: 148
• 34 HYBRIDISATION AND GENE PROBES: 167
• 36 APPLICATIONS OF GENE CLONING: 171
• 37 EXPRESSION OF FOREIGN GENES: 178
• 38 ANALYSING GENES AND GENE EXPRESSION: 184
• 39 ANALYSING WHOLE GENOMES: 196
• 310 MOLECULAR BIOTECHNOLOGY AND APPLICATIONS: 202
• 311 KEY TERMS: 204
• 312 SUGGESTIONS FOR FURTHER READING: 205
• IMMUNOCHEMICAL TECHNIQUES: 206
• 42 PRODUCTION OF ANTIBODIES: 211
• 43 PURIFICATION AND FRAGMENTATION OF IMMUNOGLOBULINS: 222
• 44 IMMUNOPRECIPITATION: 229
• 45 LABELLING ANTIBODIES: 234
• 46 IMMUNOBLOTTING: 240
• 47 IMMUNOASSAYS: 244
• 48 IMMUNOHISTOCYTOCHEMISTRY: 254
• 49 AFFINITY AND AVIDITY: 260
• 411 KEY TERMS: 261
• 412 CALCULATION: 262
• CENTRIFUGATION TECHNIQUES: 263
• 52 BASIC PRINCIPLES OF SEDIMENTATION: 264
• 53 CENTRIFUGES AND THEIR USE: 271
• 54 DESIGN AND CARE OF PREPARATIVE ROTORS: 276
• 55 SAMPLE CONTAINERS: 284
• 56 SEPARATION METHODS IN PREPARATIVE ULTRACENTRIFUGES: 285
• 57 PERFORMING DENSITY GRADIENT SEPARATIONS: 290
• 58 SELECTION EFFICIENCY AND APPLICATIONS OF PREPARATIVE ROTORS: 296
• 59 ANALYSIS OF SUBCELLULAR FRACTIONS: 300
• 510 SOME APPLICATIONS OF THE ANALYTICAL ULTRACENTRIFUGE: 302
• 511 SAFETY ASPECTS IN THE USE OF CENTRIFUGES: 305
• 512 KEY TERMS: 306
• 514 SUGGESTIONS FOR FURTHER READING: 311
• PROTEIN STRUCTURE PURIFICATION AND CHARACTERISATION: 312
• 62 PROTEIN STRUCTURE: 316
• 63 PROTEIN PURIFICATION: 318
• 64 PROTEIN STRUCTURE DETERMINATION: 338
• 65 KEY TERMS: 353
• 66 CALCULATIONS: 354
• 67 SUGGESTIONS FOR FURTHER READING: 356
• BIOMOLECULAR INTERACTIONS I ENZYMES: 357
• CHARACTERISTICS AND NOMENCLATURE: 358
• 73 ENZYME STEADYSTATE KINETICS: 359
• 74 ENZYME ASSAYS: 378
• 75 SUBSTRATE ASSAYS: 385
• 76 ENZYME PRESTEADYSTATE KINETICS: 386
• 77 ENZYME ACTIVE SITES AND CATALYTIC MECHANISMS: 389
• 78 IMMOBILISED ENZYMES: 394
• 79 CELLULAR CONTROL OF METABOLIC ACTIVITY: 396
• 710 KEY TERMS: 400
• 711 CALCULATIONS: 401
• 712 SUGGESTIONS FOR FURTHER READING: 402
• BIOMOLECULAR INTERACTIONS II CELL SURFACE RECEPTORS AND TRANSPORTERS: 403
• 82 QUANTITATIVE ASPECTS OF RECEPTORLIGAND BINDING: 406
• 83 RECEPTOR STRUCTURES: 417
• 84 MECHANISMS OF SIGNAL TRANSOUCTION: 420
• 85 SIGNAL AMPLIFICATION: 430
• 86 KEY TERMS: 432
• 88 PHYSICAL DIFFUSION: 433
• 89 FACILITATED TRANSPORT: 435
• 810 ACTIVE TRANSPORT AND ION CHANNELS: 438
• 811 RECEPTORMEDIATED ENDOCYTOSIS: 445
• 812 KEY TERMS: 449
• 814 SUGGESTIONS FOR FURTHER READING: 451
• SPECTROSCOPIC TECHNIQUES I ATOMIC AND MOLECULAR ELECTRONIC SPECTROSCOPY: 453
• 92 yRAY SPECTROSCOPY AND yRAY RESONANCE SPECTROSCOPY: 456
• 93 XRAY SPECTROSCOPY: 458
• 94 ULTRAVIOLET AND VISIBLE LIGHT SPECTROSCOPY: 459
• 95 SPECTROFLUORIMETRY: 471
• 96 CIRCULAR DICHROISM SPECTROSCOPY: 479
• 97 TURBIDIMETRY AND NEPHELOMETRY: 482
• 98 LUMINOMETRY: 483
• 99 ATOMIC SPECTROSCOPY: 485
• 910 LASERS: 490
• 911 KEY TERMS: 491
• 912 CALCULATIONS: 492
• 913 SUGGESTIONS FOR FURTHER READING: 497
• SPECTROSCOPIC TECHNIQUES II VIBRATIONAL SPECTROSCOPY AND ELECTRON AND NUCLEAR SPIN ORIENTATION IN MAGNETIC ...: 498
• 103 ELECTRON SPIN RESONANCE SPECTROSCOPY: 501
• 104 NUCLEAR MAGNETIC RESONANCE SPECTROSCOPY: 508
• 105 KEY TERMS: 525
• 106 SUGGESTIONS FOR FURTHER READING: 526
• MASS SPECTROMETRIC TECHNIQUES: 527
• 113 ELECTRON IMPACT IONISATION: 529
• 114 CHEMICAL IONISATION: 538
• 115 FIELD IONISATION: 539
• 117 ION EVAPORATION METHODS: 551
• 118 ANALYSERS: 555
• 119 DETECTORS: 564
• 1110 TANDEM MASS SPECTROMETRY: 566
• 1111 KEY TERMS: 573
• 1113 SUGGESTIONS FOR FURTHER READING: 579
• ELECTROPHORETIC TECHNIQUES: 580
• 122 SUPPORT MEDIA: 584
• 123 ELECTROPHORESIS OF PROTEINS: 588
• 124 ELECTROPHORESIS OF NUCLEIC ACIDS: 607
• 125 CAPILLARY ELECTROPHORESIS: 612
• 126 KEY TERMS: 617
• 127 CALCULATION: 618
• CHROMATOGRAPHIC TECHNIQUES: 619
• 132 CHROMATOGRAPHY THEORY AND PRACTICE: 623
• 133 LOW PRESSURE COLUMN CHROMATOGRAPHY: 631
• 134 HIGH PERFORMANCE LIQUID CHROMATOGRAPHY: 637
• 135 ADSORPTION CHROMATOGRAPHY: 647
• 136 PARTITION CHROMATOGRAPHY: 649
• 137 IONEXCHANGE CHROMATOGRAPHY: 656
• 138 MOLECULAR EXCLUSION PERMEATION CHROMATOGRAPHY: 661
• 139 AFFINITY CHROMATOGRAPHY: 665
• 1310 GASLIQUID CHROMATOGRAPHY: 672
• 1311 THINLAYER PLANAR CHROMATOGRAPHY: 678
• 1312 SELECTION OF A CHROMATOGRAPHIC SYSTEM: 681
• 1313 KEY TERMS: 682
• 1314 CALCULATIONS: 683
• 1315 SUGGESTIONS FOR FURTHER READING: 686
• RADIOISOTOPE TECHNIQUES: 687
• 142 DETECTION AND MEASUREMENT OF RADIOACTIVITY: 693
• 143 OTHER PRACTICAL ASPECTS OF COUNTING RADIOACTIVITY AND ANALYSIS OF DATA: 713
• 144 INHERENT ADVANTAGES AND RESTRICTIONS OF RADIOTRACER EXPERIMENTS: 717
• 145 SAFETY ASPECTS: 718
• 146 APPLICATIONS OF RADIOISOTOPES IN THE BIOLOGICAL SCIENCES: 721
• 147 KEY TERMS: 726
• 149 SUGGESTIONS FOR FURTHER READING: 728
• ELECTROCHEMICAL TECHNIQUES: 729
• 152 PRINCIPLES OF ELECTROCHEMICAL TECHNIQUES: 734
• 153 REDOX REACTIONS: 742
• 154 THE pH ELECTRODE: 745
• 155 IONSELECTIVE AND GASSENSING ELECTRODES: 748
• 156 THE CLARK OXYGEN ELECTRODE: 750
• 157 ELECTROCHEMICAL DETECTORS FOR HPLC: 757
• 158 BIOSENSORS: 760
• 159 KEY TERMS: 768
• 1511 SUGGESTIONS FOR FURTHER READING: 770
• INDEX: 771

Bibliographic information

• Title: Principles and Techniques of Practical Biochemistry
• Editors: Keith Wilson, John Walker
• Edition: illustrated, revised
• Publisher: Cambridge University Press, 2000
• ISBN: 052165873X, 9780521658737
• Length: 784 pages
• Subjects: Medical / Biochemistry; Science / Life Sciences / Biochemistry; Science / Scientific Instruments