Molecular Cloning: A Laboratory Manual, Volumen1

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CSHL Press, 2001 - 2344 páginas

The first two editions of this manual have been mainstays of molecular biology for nearly twenty years, with an unrivalled reputation for reliability, accuracy, and clarity.
In this new edition, authors Joseph Sambrook and David Russell have completely updated the book, revising every protocol and adding a mass of new material, to broaden its scope and maintain its unbeatable value for studies in genetics, molecular cell biology, developmental biology, microbiology, neuroscience, and immunology.
Handsomely redesigned and presented in new bindings of proven durability, this three–volume work is essential for everyone using today’s biomolecular techniques.
The opening chapters describe essential techniques, some well–established, some new, that are used every day in the best laboratories for isolating, analyzing and cloning DNA molecules, both large and small.
These are followed by chapters on cDNA cloning and exon trapping, amplification of DNA, generation and use of nucleic acid probes, mutagenesis, and DNA sequencing.
The concluding chapters deal with methods to screen expression libraries, express cloned genes in both prokaryotes and eukaryotic cells, analyze transcripts and proteins, and detect protein–protein interactions.
The Appendix is a compendium of reagents, vectors, media, technical suppliers, kits, electronic resources and other essential information.
As in earlier editions, this is the only manual that explains how to achieve success in cloning and provides a wealth of information about why techniques work, how they were first developed, and how they have evolved.

 

Contenido

Chapter 12
12
Chapter 9
11
1
13
In Vitro Amplification of DNA by the Polymerase 8 1
8
Preparation of Radiolabeled DNA and RNA Probes 9 1
9
INTRODUCTION
3
Chapter 4
4
PROTOCOLS
16

INTRODUCTION
16-13
Calciumphosphatemediated Transfection of Cells with Highmolecularweight 16 21
16-21
DNA Transfection Using Polybrene 16 43
16-43
Chapter 1
17-1
Chapter 17
17-3
Gel Electrophoresis of DNA and Pulsedfield Agarose 5 1
17-5
Transcriptional Runon Assays 17 23
17-23
INFORMATION PANELS
17-75
INFORMATION PANELS
17-91
INFORMATION PANELS
17-99
Chapter 18
18-3
Extraction Purification and Analysis of mRNA from 7 1
18-7
Chapter 10
18-10
Chapter 14
18-14
INFORMATION PANELS
18-82
INFORMATION PANELS
18-107
INFORMATION PANELS
18-115
Appendices
18-137
Chapter 13
13
1
30
INFORMATION PANELS
6
Chapter 8
8
Working with Highcapacity Vectors 4 1
4
INTRODUCTION
10
DNA Sequencing 12 1
12
Working with Synthetic Oligonucleotide Probes 10 1
10
INTRODUCTION
R-20
1
i
PREFACE XV
xv
Chapter 2
2
Screening Expression Libraries 14 1
14
1
56
Chapter 4
59
7
75
11
84
INFORMATION PANELS
109
Amplifiers
144
AnalogtoDigital Converters
155
References
165
CALIBRATION AND RESPONSE 93
173
577
183
59599
203
Derechos de autor

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Página R-7 - MJ: The early stages of absorption of injected horseradish peroxidase in the proximal tubules of mouse kidney: ultrastructural cytochemistry by a new technique.
Página 21 - The candela is the luminous intensity, in a given direction, of a source that emits monochromatic radiation of frequency 540 x 1012 hertz and that has a radiant intensity in that direction of (1/683) watt per steradian.
Página 17-5 - Science is built up with facts, as a house is with stones. But a collection of facts is no more a science than a heap of stones is a house.
Página 15-61 - Chirgwin, JM, Przybyla, AE, MacDonald, RJ, and Rutter, WJ (1979) Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18, 5294-5299.
Página R-7 - Harlow E. and Lane D. 1988. Antibodies: A laboratory manual Cold Spring Harbor Laboratory, Cold Spring Harbor, New York.
Página R-16 - Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase.
Página 15-65 - Yanisch-Perron, C., Vieira, J., and Messing, J. (1985). Improved M13 Phage Cloning Vectors and Host Strains: Nucleotide Sequences of the M13mpl8 and PUC19 Vectors.
Página 15-65 - Smith, DB, and Johnson, KS (1988). Single-step purification of polypeptides expressed in Escherichia coli as fusions with glutathione S-transferase. Gene 67, 31-40.
Página 17-1 - Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter.

Información bibliográfica