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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.
Part 1 Essentials
1. Isolation and Quantification of DNA
2. Analysis of DNA
3. Cloning and Transformation with Plasmid Vectors
4. Gateway Recombinational Cloning
5. Working with Bacterial Artificial Chromosomes and Other High-Capacity Vectors
6. Extraction, Purification, and Analysis of RNA from Eukaryotic Cells
7. Polymerase Chain Reaction
Part 2 Analysis and Manipulations of DNA AND RNA
9. Quantification of DNA and RNA by Real-Time Polymerase Chain Reaction
10. Nucleic Acid Platform Technologies
11. DNA Sequencing
12. Analysis of DNA Methylation in Mammalian Cells
13. Preparation of Labeled DNA, RNA, and Oligonucleotide Probes
14. Methods for In Vitro Mutagenesis
Part 3 Introducing Genes into Cells
15. Introducing Genes into Cultured Mammalian Cells
16. Introducing Genes into Mammalian Cells: Viral Vectors
Part 4 Gene Expression
17. Analysis of Gene Regulation Using Reporter Systems
18. RNA Interference and Small RNA Analysis
19. Expressing Cloned Genes for Protein Production, Purification, and Analysis
Part 5 Interaction Analysis
20. Cross-Linking Technologies for Analysis of Chromatin Structure and Function
21. Mapping of In Vivo RNA-Binding Sites by UV-Cross-Linking Immunoprecipitation (CLIP)
22. Gateway-Compatible Yeast One-Hybrid and Two-Hybrid Assays
1. Reagents and Buffers
2. Commonly Used Techniques
3. Detection Systems
4. General Safety and Hazardous Material
"In science [...] speed matters and credibility matters. And compared to banking on [...] an anonymous contributor in an online discussion group, you'd be better off trusting Michael Green and Joseph Sambrook. When skimming through the pages of Molecular Cloning, it is impossible not to gain new insights that will push your experiments forward."
– Lab Times
Praise for the previous edition:
"Any basic research laboratory using molecular biology techniques will benefit from having a copy on hand of the newly published Third Edition of Molecular Cloning: A Laboratory Manual [...] the first two editions of this book have been staples of molecular biology with a proven reputation for accuracy and thoroughness."
– The Scientist
"In every kitchen there is at least one indispensable cookbook [...] Molecular Cloning: A Laboratory Manual fills the same niche in the laboratory (with) information to help both the inexperienced and the advanced user. (It) has once again established its primacy as the molecular laboratory manual and is likely to be found on lab benches [...] around the world."
– Trends in Neurosciences
"Molecular Cloning: A Laboratory Manual has always been the laboratory mainstay for protocols and techniques. It has a pure-bred ancestry, and the new edition does not disappoint. (It) includes information panels at the end of each chapter that describe the principles behind the protocols [...] The addition of this information extends Molecular Cloning from an essential laboratory resource into a new realm, one merging the previous prototype with a modern molecular monograph [...] the next generation of Molecular Cloning not only carries on the proud heritage of the first two editions but also admirably expands on that tradition to provide a truly essential laboratory manual."
– Trends in Microbiology