Principles and Techniques of Practical Biochemistry

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

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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
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

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
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
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About the author (2000)

Keith Wilson is professor and former chair of English at the University of Ottawa and Honorary Vice President of the Thomas Hardy Association.

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