Molecular Cloning: A Laboratory Manual (3-Volume Set)

by Joseph Sambrook, David W. Russell, Joe Sambrook

Publisher: Cold Spring Harbor Laboratory
ISBN: 1936113422

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Book Description

Molecular Cloning has served as the foundation of technical expertise in labs worldwide for 30 years. No other manual has been so popular, or so influential. Molecular Cloning, Fourth Edition, by the celebrated founding author Joe Sambrook and new co-author, the distinguished HHMI investigator Michael Green, preserves the highly praised detail and clarity of previous editions and includes specific chapters and protocols commissioned for the book from expert practitioners at Yale, U Mass, Rockefeller University, Texas Tech, Cold Spring Harbor Laboratory, Washington University, and other leading institutions.

The theoretical and historical underpinnings of techniques are prominent features of the presentation throughout, information that does much to help trouble-shoot experimental problems. For the fourth edition of this classic work, the content has been entirely recast to include nucleic-acid based methods selected as the most widely used and valuable in molecular and cellular biology laboratories. Core chapters from the third edition have been revised to feature current strategies and approaches to the preparation and cloning of nucleic acids, gene transfer, and expression analysis.

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 Joe 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.

Table of Contents

1. Plasmids and Their Usefulness in Molecular Cloning
2. Bacteriophage lambda and Its Vectors
3. Working with Bacteriophage M13 Vectors
4. Working with High-capacity Vectors
5. Gel Electrophoresis of DNA and Pulsed-field Agarose Gel Electrophoresis
6. Preparation and Analysis of Eukaryotic Genomic DNA
7. Extraction, Purification, and Analysis of mRNA from Eukaryotic Cells
8. In vitro Amplification of DNA by the Polymerase Chain Reaction
9. Preparation of Radiolabeled DNA and RNA Probes
10. Working with Synthetic Oligonucleotide Probes
11. Preparation of cDNA Libraries and Gene Identification
12. DNA Sequencing
13. Mutagenesis
14. Screening Expression Libraries
15. Expression of Cloned Genes in Escherichia coli
16. Introducing Cloned Genes into Cultured Mammalian Cells
17. Analysis of Gene Expression in Cultured Mammalian Cells
18. Analysis of Protein-protein Interactions

Customer Reviews

In this 3 volume set of books the authors summarize the most important laboratory protocols for DNA analysis and cloning. As someone involved in computational biology and mathematical gene sequence analysis, I was needing such a summary to get an idea of just how genetic engineering is actually practiced in the laboratory. The book is definitely written for those readers that are very experienced in these "wet" techniques, but it still could be perused profitably by anyone who is curious about genetic engineering. There is also an excellent website that owners of the books can go to and search for protocols and obtain updates and additions to the protocols.

At the beginning of each chapter, the authors give an introduction to the protocols and this is of an enormous help to those readers with only rudimentary acquaintance with the laboratory procedures. Typically, this introduction contains an historical summary of the procedures as they were developed or discovered. One can only marvel at the ingenuity of the discoverers of these techniques. These introductions are fairly straightforward to read, even for those that are not experts in biochemistry.

At the end of each chapter, the authors include an "information panel" that gives a more in-depth view of the biochemistry or genetics behind the procedures. These are summaries and are highly specialized, and are again meant for experienced readers. A very lengthy list of references is also included at the end of each chapter.

Becuase of the size of this collection, space here does not permit a detailed review, so I will list some of the areas that I thought were particularly interesting or well-written (these coming from the introduction or the information panels only):

1. The DNA synthesis at the colE1 replicon and the interaction between RNAI and RNAII.
2. The discussion of electroporation and the physics behind this technique to introduce DNA into eukaryotic cells.
3. The discussion on the discovery of bacteriophage lambda.
4. The discussion (with diagram), of the assembly pathway of bacteriophage lambda.
5. The summary of the early analysis of DNA using electrophoresis and the different pulsed-field configurations used.
6. The anecdote on the discovery of the polymerase chain reaction.
7. The short discussion on computer-assisted design of oligonucleotide primers.
8. The discussion of oligonucleotide synthesis.
9. The flowchart detailing the preparing and screening of a cDNA library.
10. The history of the development of the methods to synthesize and clone cDNAs.
11. The detailed discussion of the molecular cloning of double-stranded cDNA.
12. The discussion on the methods to validate clones of cDNA.
13. The discussion on magnetic beads for affinity purification.
14. The discussion on the history of DNA sequencing and the different techniques to accomplish it, particularly the information panel on automated DNA sequencing.
15. The discussion of the different types of mutagenesis and the different methods for accomplishing it.
16. The fascinating discussion of how to introduce cloned genes into mammalian cells.
17. The discussion on the steps involved in DNA footprinting.
18 The discussion on green flourescent protein and its use as a fusion tag.
19. The discussion on the use of surface plasmon resonance spectroscopy.

I love this book, however some areas are better covered in Current Protocols in Molecular Biology, and vice versa. So I would recommend having both.

Any lab serious about molecular biology has this book. A lab staple for over 20 years the new updated version is even more comprehensive reflecting the continuing development of molecular biology. Most importantly the series still retains the historical significance and the background or reference material for many of the techniques. A must have!

A classic reference work in Molecular Biology, newly and completely updated. It has extensive coverage of nearly all core molecular biology techniques and often presents several different means of accomplishing the same goal. The website is easy to access and the protocols are clearly written. The website makes it easy to print out protocols. In addition each protocol contains references to the primary literature supporting its claims. In the internet version of the protocol the references are hotlinked to PubMed, greatly simplifying access to the underlying references. I definitely recommend this book to all labs using molecular biology. Make sure to check out the website!

Sambrook and Russell answer every question you could think of, and then some. This book is a goldmine of information, packed with protocols, but also filled with the extra information that transforms a simple set of instructions into an amazingly helpful how-to manual. The approach taken is that of an experimentor (ie. 'How to win the battle with RNase'--it really is a battle!), with hints and suggestions usually learned by watching an old pro. The information is well-organized, and very well illustrated to give a clear view of how an experiment is performed and especially the logic behind it. The manual also satisfies the insatiable curiosity of a scientist rather than a technician: how do pharmacological agents work? What are their structures? And the troubleshooting sections provide an invaluable resource. All in all, Sambrook and Russell have created an essential weapon in any scientist's artillery.

Molecular cloning has been a lab staple for years. Now reprinted so you can update the old lab copy worn out by years of student use! Its a must have for any lab serious about molecular biology. Its also useful for student training. Many times there are simple explanations for the lab techniques we have adopted as dogma, but are unsure why. Molecular cloning has the answers and is a great resource. I highly recommend this book for its depth and breadth of protocols and guidance in the complicated realm of cloning!

This book is an invaluable resource for any serious practitioner of molecular biology. Content is detailed and comprehensive. Highly recommended reference source. All credit to the authors for what is a thorough revision of this latest testament of what is undoubtedly the revealed scripture of molecular biology.

You should read the upgraded version. This version lacks things but also explains alot. If you practice molecular biology you should keep this book near

In the lab where I work we have this volume. We were required to read certain parts if it. I liked how the books explained complicated parts of molecular cloning. The bad part the books are very long.

This set of three manuals is invaluable in the lab. Complete with almost every technique commonly used, and full useful tables and other data sets that save the user a lot of time. One key point though, is that a new edition is long overdue.