This volume contains a comprehensive compilation of chromogenic and fluorescent RNA in situ hybridization (ISH) technology in many of its various shades, forms, and applications. The book is organized into a number of parts and chapters focusing on the application of ISH methodologies to different animal species as used in Evolutionary Development (EvoDevo) and Biomedical research, and covering new developments in RNA visualization by fluorescent ISH (FISH). The described (F)ISH protocols employ effective strategies for signal enhancement and target amplification allowing for high signal intensities and drastically improved signal-to-noise ratios. Chromogenic and fluorescent ISH, as specified in the various chapters, are most essential for RNA expression profiling, applied to many fields of research including cellular, developmental, and evolutionary biology, neurobiology and neuropathology. Written for the popular Neuromethods series, chapters include the kind of detail and key implementation advice that ensures successful results in the laboratory. Essential and authoritative, In Situ Hybridization Methods provides detailed protocols for newcomers to ISH, and inspires researchers familiar with the technique to seek and find up-to-date methodology for new and specialized applications.

In Situ Hybridization Protocols, Fourth Edition contains 21 protocols that utilize the in situ hybridization technology to document or take advantage of the visualization of specific RNA molecules. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, In Situ Hybridization Protocols, Fourth Edition seeks to aid scientists in the further discovery of new RNA species and uncovering of their cellular functions.

This book is a unique source of information on the present state of the exciting field of molecular cytogenetics and how it can be applied in research and diagnostics. The basic techniques of fluorescence in situ hybridization and primed in situ hybridization (PRINS) are outlined, the multiple approaches and probe sets that are now available for these techniques are described, and applications of them are presented in 36 chapters by authors from ten different countries around the world. The book not only provides the reader with basic and background knowledge on the topic, but also gives detailed protocols that show how molecular cytogenetics is currently performed by specialists in this field. The FISH Application Guide initially provides an overview of the (historical) development of molecular cytogenetics, its basic procedures, the equipment required, and probe generation. The book then describes tips and tricks for making different tissues available for molecular cytogenetic studies. These are followed by chapters on various multicolor FISH probe sets, their availability, and their pot- tial for use in combination with other approaches. The possible applications that are shown encompass the characterization of marker chromosomes, cryptic cytogenetic aberrations and epigenetic changes in humans by interphase and metaphase cyto- netics, studies of nuclear architecture, as well as the application of molecular cytogenetics to zoology, botany and microbiology.

Annotation Darby (human biology, RMIT U., Victoria, Australia) is joined by geneticists, molecular biologists, and pathologists from around the world to describe basic and advanced techniques for hybridization, for whole-mount embryo specimens and at the electron microscope level. Coverage includes protocols for detection of DNA fragmentation in apoptosis, localization of genes to particular chromosomes, and the use of DNA and RNA probes to detect expression in cells or tissue sections. For novice and experienced investigators who need proven and readily reproducible methods. Annotation c. Book News, Inc., Portland, OR (booknews.com)

Molecular biology and genetic engineering techniques have revolutionized our concepts of gene activity in cellular function. In situ hybridization has contributed to this revolution. It is the only technique which gives precise data on the content and distribution of DNA and mRNA in intactcells and tissues. It is now an invaluable test in understanding the pathophysiology of cellular functions. This book covers all aspects of in situ hybridization. It describes practical procedures and protocols, the scientific background, areas of application, and the limitations of the technology. The contributing authors are all experts. Each chapter contains up-to-date references, many colourillustrations, the latest advances, and prospects for the future. This practical guide will be invaluable to: molecular, cell, and developmental biologists; cyto- and molecular geneticists; clinicians, especially pathologists, biologists, and microbiologists; and scientists seriously interested inthe pathophysiology of cell function and its regulation.

In situ hybridization is used to reveal the location of specific nucleic acids sequences on chromosomes or in tissues. Visualization of the location of genes on chromosomes or of specific mRNAs or viruses in tissues is crucial for understanding the organization, regulation, and function of genes. It is a therefore a core technique in all areas of biomedical research. In Situ Hybridization: A Practical Approach 2/e is the second edition of one of the most successful Practical Approach books, published in 1992. Since the first edition was published, a number of important technical advances have been made. The new edition has been thoroughly updated to contain protocols detailing the major techniques of in situ hybridization currently in use: in situ hybridization to mRNA with oligonucleotide and RNA probes (radiolabelled and hapten labelled); analysis using light and electron microscopes; whole mount in situ hybridization; double detection of RNAs, and RNA plus protein; and fluorescent in situ hybridization to detect chromosomal sequences. The protocols are complemented by advice on strategies for successful results, descriptions of the theoretical basis of in situ hybridization and important new developments in gene expression databases. The procedures described are widely applicable to many systems. The use of in situ hybridization in PCR is covered in a separate volume: Herrington and O'Leary (Eds) PCR 3 - PCR in situ hybridization: A Practical Approach (OUP, 1997). All the authors have extensive practical experience of establishing reliable techniques of in situ hybridization. This book will be useful to all researchers at all levels who use in situ hybridization.