Principles and Techniques of Biochemistry and Molecular Biology

by Edited by Keith Wilson , John Walker

• ISBN13: 9780521535816
• ISBN10: 0521535816
• Edition: 6th
• Format: Paperback
• Copyright: 2005-03-21
• Publisher: Cambridge University Press

Summary

This is a new and expanded edition of the bestselling Principles and Techniques of Practical Biochemistry. It includes discussion of the theoretical principles, practical details and applications of the key experimental techniques that are routinely used in modern biochemistry and molecular biology, including those that have led to the emergence of the new disciplines of genomics, proteomics and bioinformatics. All chapters have been updated and new sections added to cover the principles of experimental design, the statistical analysis of quantitative analytical data, and the principles and practice of clinical biochemistry. here are new chapters covering cell culture, microscopy and mass spectrometry. To enhance student understanding of each topic, in-text worked examples are included in each chapter. The textbook will be essential reading for all bioscience students and pre-clinical medical students.

Table of Contents

Preface to the sixth edition	xi
List of contributors	xiii
List of abbreviations	xvi
Basic principles	1	(70)
K. Wilson
J. Fyffe
Biochemical studies	1	(2)
Units of measurements	3	(7)
Weak electrolytes	10	(5)
Buffer solutions-their nature and preparation	15	(3)
pH and oxygen electrodes	18	(10)
Quantitative biochemical measurements	28	(20)
Principles of clinical biochemical analysis	48	(20)
Safety in the laboratory	68	(1)
Suggestions for further reading	69	(2)
Cell culture techniques	71	(32)
A. R. Baydoun
Introduction	71	(1)
The cell culture laboratory and equipment	72	(4)
Safety considerations in cell culture	76	(1)
Aseptic techniques and good cell culture practice	77	(4)
Types of animal cell and their characteristics in culture	81	(12)
Bacterial cell culture	93	(4)
Plant cell culture	97	(4)
Potential use of cell cultures	101	(1)
Suggestions for further reading	101	(2)
Centrifugation	103	(28)
K. Ohlendieck
Introduction	103	(1)
Basic principles of sedimentation	104	(5)
Types, care and safety aspects of centrifuges	109	(8)
Preparative centrifugation	117	(7)
Analytical centrifugation	124	(6)
Suggestions for further reading	130	(1)
Microscopy	131	(35)
S. W. Paddock
Introduction	131	(2)
The light microscope	133	(13)
Optical sectioning	146	(5)
Imaging living cells and tissues	151	(2)
The stereomicroscope	153	(1)
The electron microscope	154	(5)
Imaging and biochemistry	159	(1)
Specialised imaging techniques	160	(2)
Image archiving, presentation and further information	162	(1)
Suggestions for further reading	163	(3)
Molecular biology, bioinformatics and basic techniques	166	(126)
R. Rapley
Introduction	166	(1)
Structure of nucleic acids	167	(7)
Genes and genome complexity	174	(4)
Location and packaging of nucleic acids	178	(1)
Functions of nucleic acids	179	(11)
The manipulation of nucleic acids: basic tools and techniques	190	(1)
Isolation and separation of nucleic acids	191	(7)
Molecular biology and bioinformatics	198	(2)
Molecular analysis of nucleic acid sequences	200	(7)
The polymerase chain reaction	207	(9)
Nucleotide sequencing of DNA	216	(8)
Suggestions for further reading	224	(1)
Recombinant DNA and genetic analysis
R. Rapley	225	(1)
Introduction	225	(1)
Constructing gene libraries	225	(10)
Cloning vectors	235	(18)
Hybridisation and gene probes	253	(2)
Screening gene libraries	255	(4)
Applications of gene cloning	259	(5)
Expression of foreign genes	264	(6)
Analysing genes and gene expression	270	(13)
Analysing whole genomes	283	(5)
Pharmacogenomics	288	(1)
Molecular biotechnology and its applications	289	(2)
Suggestions for further reading	291	(1)
Immunochemical techniques	292	(57)
R. Thorpe
S. Thorpe
Introduction	292	(6)
Production of antibodies	298	(10)
Purification and fragmentation of immunoglobulins	308	(7)
Immunoprecipitation	315	(5)
Labelling antibodies	320	(8)
Immunoblotting	328	(3)
Immunoassays	331	(10)
Immunohisto/cytochemistry	341	(6)
Affinity and avidity	347	(1)
Immunochemical use of surface plasmon resonance	347	(1)
Suggestions for further reading	348	(1)
Protein structure, purification, characterisation and function analysis	349	(56)
J. M. Walker
Ionic properties of amino acids and proteins	349	(4)
Protein structure	353	(3)
Protein purification	356	(22)
Protein structure determination	378	(14)
Proteomics and protein function	392	(12)
Suggestions for further reading	404	(1)
Mass spectrometric techniques	405	(44)
A. Aitken
Introduction	405	(2)
Ionisation	407	(6)
Mass analysers	413	(17)
Detectors	430	(1)
Structural information by tandem mass spectrometry	430	(13)
Analysing protein complexes	443	(1)
Computing and database analysis	444	(3)
Suggestions for further reading	447	(2)
Electrophoretic techniques	449	(36)
J. M. Walker
General principles	449	(4)
Support media	453	(4)
Electrophoresis of proteins	457	(16)
Electrophoresis of nucleic acids	473	(5)
Capillary electrophoresis	478	(5)
Microchip electrophoresis	483	(1)
Suggestions for further reading	484	(1)
Chromatographic techniques	485	(66)
K. Wilson
Principles of chromatography	485	(4)
Chromatographic performance parameters	489	(11)
Liquid chromatography (LPLC and HPLC)	500	(15)
Adsorption chromatography	515	(3)
Partition chromatography	518	(6)
Ion-exchange chromatography	524	(5)
Molecular exclusion (gel filtration) chromatography	529	(4)
Affinity chromatography	533	(8)
Gas-liquid chromatography	541	(5)
Thin-layer (planar) chromatography	546	(3)
Selection of a chromatographic system	549	(1)
Suggestions for further reading	550	(1)
Spectroscopic techniques: I Atomic and molecular electronic spectroscopy	551	(42)
D. B. Gordon
Introduction	551	(3)
γRay spectroscopy and γ-ray resonance spectroscopy	554	(2)
X-ray spectroscopy	556	(1)
Ultraviolet and visible light spectroscopy	557	(14)
Spectrofluorimetry	571	(8)
Circular dichroism spectroscopy	579	(4)
Turbidimetry and nephelometry	583	(1)
Luminometry	584	(2)
Atomic spectroscopy	586	(5)
Lasers	591	(1)
Suggestions for further reading	592	(1)
Spectroscopic techniques: II Vibrational spectroscopy and electron and nuclear spin orientation in magnetic fields	593	(28)
D. B. Gordon
Introduction	593	(1)
Infrared and Raman spectroscopy	594	(2)
Electron spin resonance spectroscopy	596	(7)
Nuclear magnetic resonance spectroscopy	603	(17)
Suggestions for further reading	620	(1)
Radioisotope techniques	621	(44)
R. J. Slater
The nature of radioactivity	621	(7)
Detection and measurement of radioactivity	628	(23)
Other practical aspects of counting radioactivity and analysis of data	651	(4)
Inherent advantages and restrictions of radiotracer experiments	655	(1)
Safety aspects	656	(3)
Applications of radioisotopes in the biological sciences	659	(5)
Suggestions for further reading	664	(1)
Enzymes	665	(54)
K. Wilson
Characteristics and nomenclature	665	(3)
Analytical methods for the study of enzyme reactions	668	(11)
Enzyme steady-state kinetics	679	(23)
Enzyme active sites and catalytic mechanisms	702	(7)
Control of enzyme activity	709	(9)
Suggestions for further reading	718	(1)
Cell membrane receptors	719	(50)
K. Wilson
Receptors for cell signalling	719	(1)
Quantitative aspects of receptor-ligand binding	720	(9)
Techniques for the study of receptor-ligand binding	729	(17)
Molecular structure of receptors	746	(5)
Mechanisms of signal transduction	751	(12)
Receptor desensitisation and trafficking	763	(4)
Suggestions for further reading	767	(2)
Index	769
The colour figure section is between pp. 142 and 143