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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.

They are augmented by 12 new chapters which show how DNA, RNA, and

proteins should be prepared, evaluated, and manipulated, and how

data generation and analysis can be handled.

The new content includes methods for studying interactions

between cellular components, such as microarrays, next-generation

sequencing technologies, RNA interference, and epigenetic analysis

using DNA methylation techniques and chromatin immunoprecipitation.

To make sense of the wealth of data produced by these techniques, a

bioinformatics chapter describes the use of analytical tools for

comparing sequences of genes and proteins and identifying common

expression patterns among sets of genes.

Building on thirty years of trust, reliability, and authority,

the fourth edition of Molecular Cloning is the new gold

standard—the one indispensable molecular biology laboratory manual

and reference source.

JOSEPH SAMBROOK：

Sambrook was educated at the University of Liverpool (BSc (hons)

1962) and obtained his PhD at the Australian National University in

1966. He did postdoctoral research at the MRC Laboratory of

Molecular Biology (1966-67) and the Salk Institute for Biological

Studies (1967-69). In 1969 he was hired by James D. Watson to work

at the Cold Spring Harbor Laboratory in New York. Watson has been

reported to say this was the best hiring decision he ever made. Joe

was responsible for creating a combative creative environment at

CSHL that fomented discovery. Subsequently he worked at the

University of Texas Southwestern Medical Center (Dallas).

Sambrook is best known for his studies on DNA tumor viruses and

the molecular biology of normal and neoplastic cells. His Tumour

Virus Group at Cold Spring Harbor identified and mapped all of the

major genes of adenoviruses and SV40, determined their

transcriptional control in infected and transformed cells, and

elucidated the mechanism of integration of these viruses into the

genome of the host cell.[citation needed] He has also made

important contributions to the understanding of intracellular

traffic and protein folding and is an influential leader in the

field of the molecular genetics of human cancer.

VOLUME 1
Part 1: Essentials
1. Isolation and Quantification of DNA
2. Analysis of DNA
3. Cloning and Transformation with Plasmid Vectors
4. Gateway Recombinational Cloning
John S. Reece-Hoyes and Albertha J.M. Walhout
5. Working with Bacterial Artificial Chromosomes and Other
High-Capacity Vectors
Nathaniel Heintz and Shiaoching Gong
6. Extraction, Purification, and Analysis of RNA from Eukaryotic
Cells
7. Polymerase Chain Reaction
8. Bioinformatics
Jui-Hung Hung and Zhiping Weng
VOLUME 2
Part 2: Analysis and Manipulation of DNA and RNA
9. Quantification of DNA and RNA by Real-Time Polymerase Chain
Reaction
10. Nucleic Acid Platform Technologies
Oliver Rando
11. DNA Sequencing
Elaine Mardis and W. Richard McCombie
12. Analysis of DNA Methylation in Mammalian Cells
Paul M. Lizardi, Qin Yan, and Narendra Wajapeyee
13. Preparation of Labeled DNA, RNA, and Oligonucleotide
Probes
14. Methods for In Vitro Mutagenesis
Matteo Forloni, Alex Liu, and Narendra Wajapeyee
Part 3: Introducing Genes into Cells
15. Introducing Genes into Cultured Mammalian Cells
Priti Kumar, Arvindhan Nagarajan, and Pradeep D. Uchil
16. Introducing Genes into Mammalian Cells: Viral Vectors
Guangping Gao and Miguel Sena-Esteves
VOLUME 3
Part 4: Gene Expression
17. Analysis of Gene Regulation Using Reporter Systems
Pradeep D. Uchil, Arvindhan Nagarajan, and Priti Kumar
18. RNA Interference and Small RNA Analysis
Chengjian Li and Phillip D. Zamore
19. Expressing Cloned Genes for Protein Production, Purification,
and Analysis
Clara L. Kielkopf, William Bauer, and Ina Urbatsch
Part 5: Interaction Analysis
20. Cross-Linking Technologies for Analysis of Chromatin Structure
and Function
Tae Hoon Kim and Job Dekker
21. Mapping of In Vivo RNA-Binding Sites by UV-Cross-Linking
Immunoprecipitation (CLIP)
Jennifer C. Darnell, Aldo Mele, Ka Ying Sharon Hung, and Robert B.
Darnell
22. Gateway-Compatible Yeast One-Hybrid and Two-Hybrid Assays
John S. Reece-Hoyes and Albertha J.M. Walhout
Appendices
1. Reagents and Buffers
2. Commonly Used Techniques
3. Detection Systems
4. General Safety and Hazardous Material
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