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The correct procedures you need for frustration-free PCR methods and applications are contained in this complete, step-by-step, clearly written, inexpensive manual.Avoid contamination--with specific instructions on setting up your labAvoid cumbersome molecular biological techniquesDiscover new applications

Viral Vectors for Gene Therapy: Methods and Protocols consists of 30 ch- ters detailing the use of herpes viruses, adenoviruses, adeno-associated viruses, simple and complex retroviruses, including lentiviruses, and other virus systems for vector development and gene transfer. Chapter cont- butions provide perspective in the use of viral vectors for applications in the brain and in the central nervous system. Viral Vectors for Gene Therapy: Methods and Protocols contains step-by-step methods for successful rep- cation of experimental procedures, and should prove useful for both experienced investigators and newcomers in the field, including those beginning graduate study or undergoing postdoctoral training. The “Notes” section contained in each chapter provides valuable troublesho- ing guides to help develop working protocols for your laboratory. With Viral Vectors for Gene Therapy: Methods and Protocols, it has been my intent to develop a comprehensive collection of modern molecular methods for the construction, development, and use of viral vectors for gene transfer and gene therapy. I would like to thank the many chapter authors for their contributions. They are all experts in various aspects of viral vectors, and I appreciate their efforts and hard work in developing comprehensive chapters. As editor, it has been a privilege to preview the development of Viral Vectors for Gene Therapy: Methods and Protocols, and to acquire insight into the various methodological approaches from the many different contri- tors.

Superantigen Protocols assembles experimental protocols that have proved useful for the study of superantigens. These techniques will allow researchers from various areas of cell biology, microbiology, immunology, biochemistry, and molecular biology to assess the physical characteristics and biological effects of well-known superantigens as well as of putative substances that might have superantigenic activities, and to explore therapies for superantig- induced effects. Microbial exotoxins have been studied for decades as virulence factors because of their pathogenic effects. The term “superantigen” was coined by Marrack and Kappler a decade ago for some of these molecules because of their potent T-cell stimulatory activities. In recent years, advances in mole- lar biology provide recombinant as well as natural superantigens in highly purified form for physical characterization. Superantigens are now used extensively as tools to study interactions between receptors on cells of the immune system as they bind to major histocompatibility complex class II m- ecules on antigen-presenting cells and V regions of T-cell receptors. The b- ? logical effects that result from these interactions are studied both in vitro and in vivo. The intent of this book is, therefore, to bring together up-to-date te- niques developed by experts in the field of biochemistry, immunology, and molecular biology for the study of superantigens. Superantigen Protocols begins with an overview of the field to provide background information on the various classes of superantigens and their str- ture.

Using molecular methods for plant disease diagnosis provides diagnosticians with a number of advantages over more traditional methods. They can allow the identification of morphologically similar species, for example, or the detection of infection prior to symptom formation. Not only can molecular tools help by increasing the efficacy, accuracy and speed of diagnosis; their common technological basis provides further benefits, especially where resources are limited and traditional skills are hard to sustain.This book provides protocols for nucleic acid-based methods currently applied to plant pathogen detection and identification. It takes the practitioner through the full range of molecular diagnostic and detection methods, and, as these generic techniques are appropriate for use on any target with minimal modification, also provides a useful resource for students of plant pathology and plant pathologists. Beginning with the background and future directions of the science, it then addresses DNA barcoding, microarrays, polymerase chain reactions (PCR), quality assurance and more, forming a complete reference on the subject.

Homologous recombination is important in various aspects of DNA metabolism, including damage repair, replication, telomere maintenance, and meiosis, and yeast genetics has successfully provided a framework for the mechanism of homologous recombination. Divided into four convenient sections, DNA Recombination: Methods and Protocols covers recent techniques that best utilize the advantages of the yeast system, prescribing to the belief that yeast will keep serving as a great model organism to study homologous recombination. Chapters have also been included for such exceptions as the group of genes involved in recombination that are found solely in higher eukaryotes, such as BRCA2. And looking forward, a necessary step in the direction of understanding the homologous recombination process is to isolate the machine and let it work in a test tube.  Understanding the design by studying the appearance and behavior of the machinery as a single molecule will be an important milestone toward understanding the mechanism of action of the machinery. Techniques covering these topics have also been included.  Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, DNA Recombination: Methods and Protocols serves as an ideal guide to scientists of all backgrounds with its well-honed methodologies and strives to bring the reader to the next level of understanding regarding this vital subject.

A desktop companion to the three-volume Current Protocols in Molecular Biology, the recognized leader in bioscience laboratory manuals. This edition contains over 220 protocols from leading laboratories worldwide. All methods are lab-tested and include step-by-step instructions, equipment and materials necessary to successfully conduct an experiment.

Agrobacterium tumefaciens is a soil bacterium that for more than a century has been known as a pathogen causing the plant crown gall disease. Unlike many other pathogens, Agrobacterium has the ability to deliver DNA to plant cells and permanently alter the plant genome. The discovery of this unique feature 30 years ago has provided plant scientists with a powerful tool to genetically transform plants for both basic research purposes and for agric- tural development. Compared to physical transformation methods such as particle bomba- ment or electroporation, Agrobacterium-mediated DNA delivery has a number of advantages. One of the features is its propensity to generate single or a low copy number of integrated transgenes with defined ends. Integration of a single transgene copy into the plant genome is less likely to trigger “gene silencing” often associated with multiple gene insertions. When the first edition of Agrobacterium Protocols was published in 1995, only a handful of plants could be routinely transformed using Agrobacterium. Ag- bacterium-mediated transformation is now commonly used to introduce DNA into many plant species, including monocotyledon crop species that were previously considered non-hosts for Agrobacterium. Most remarkable are recent devel- ments indicating that Agrobacterium can also be used to deliver DNA to non-plant species including bacteria, fungi, and even mammalian cells.

Viral Vectors for Gene Therapy: Methods and Protocols consists of 30 ch- ters detailing the use of herpes viruses, adenoviruses, adeno-associated viruses, simple and complex retroviruses, including lentiviruses, and other virus systems for vector development and gene transfer. Chapter cont- butions provide perspective in the use of viral vectors for applications in the brain and in the central nervous system. Viral Vectors for Gene Therapy: Methods and Protocols contains step-by-step methods for successful rep- cation of experimental procedures, and should prove useful for both experienced investigators and newcomers in the field, including those beginning graduate study or undergoing postdoctoral training. The “Notes” section contained in each chapter provides valuable troublesho- ing guides to help develop working protocols for your laboratory. With Viral Vectors for Gene Therapy: Methods and Protocols, it has been my intent to develop a comprehensive collection of modern molecular methods for the construction, development, and use of viral vectors for gene transfer and gene therapy. I would like to thank the many chapter authors for their contributions. They are all experts in various aspects of viral vectors, and I appreciate their efforts and hard work in developing comprehensive chapters. As editor, it has been a privilege to preview the development of Viral Vectors for Gene Therapy: Methods and Protocols, and to acquire insight into the various methodological approaches from the many different contri- tors.

This volume seeks to enable the discovery of tools in chemical biology by providing readers with various techniques ranging from initial chemical genetic screening to target identification. To successfully highlight the essential components of the chemical biology tool discovery process, the book is organizes into four parts that focus on platforms for molecular discovery in in vitro cellular systems, in vivo chemical genetic screening protocols, and methods used to discover functional protein targets. Written in the highly successful Methods of 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 key tips on troubleshooting and avoiding known pitfalls.Practical and informative, Chemical Biology: Methods and Protocols seeks to improve the success rate of the chemical biology field through the dissemination of detailed and experiential knowledge.

The orchid family is one of the largest families of flowering plants known for their beauty and economic importance. This work provides information in key areas of research that are important to both scientists and commercial growers alike. The main purposes of this book are to provide key practical areas of research, such as, germination, micropropagation, traditional and current techniques related to plant improvement; document methods that ensure survival of plants from laboratories to greenhouses; promote communication between scientists and growers, so that their combined expertise on these areas will lead to the successful growth of orchids in their natural habitats or commercial greenhouses. This book can serve as reference for laymen with an interest in orchid growing.This book is divided into 5 parts. The first part emphasizes propagation methods using seeds and related techniques that are important to plant conservation and improvement. Successes in asymbiotic and symbiotic seed germination are keys to orchid conservation and their propagation.  The second part summarizes micropropagation methods, common media, and newer methods of micropropagation such as the bioreactor culture procedures. The third part focuses on techniques related to the manipulation of explants in an in vitro environment. The fourth part covers cell biological methods and transformation techniques. Since the successes in a laboratory setting do not guarantee plant survival and propagation in greenhouses and in the natural environment, it discusses greenhouse propagation techniques that are essential to the survival of plants generated from a laboratory setting. The fifth part showcases recent successes on orchid propagation by documenting sample publications and how to present orchids in an artistic fashion for one’s enjoyment.

John Sinclair and a panel of expert investigators present a comprehensive collection of cellular and molecular techniques for the analysis of cytomegalovirus (CMV) biology and its pathogenetic mechanisms. The methods-all described in step-by-step detail with ready reproducibility in mind-range from basic virus culture to complex molecular analysis of CMV structure and function. Included are methods for CMV detection using both immunological and biological techniques, methods for analyzing fundamental aspects of the CMV infection cycle, and methods for analyzing T cell response to cytomegalovirus infection in the human host. Comprehensive and state-of-the-art, Cytomegalovirus Protocols provides investigators with a collection of the key methods that are illuminating not only the basic biology of this complex and intriguing human herpesvirus, but also its significant role in human infectious diseases and their emergent therapies.

Drosophila is a comprehensive collection of methods and protocols for Drosophila, one of the oldest and most commonly used model organisms in modern biology. The protocols are written by the scientists who invented the methods. The text presents a diverse set of techniques that range from the basic handling of flies to more complex applications. This is the perfect reference manual for Drosophila researchers.

This fully-updated book provides systems-based methodologies and innovative technologies that can be used for solving complicated problems of complex systems. A wide range of theoretical and experimental approaches are introduced for practical applications, as the content explores the basic and novel concepts in psychoneuroimmunology (PNI), especially the relationships among stress, inflammation, and psychophysiological disorders, as well as various cutting-edge technologies and methods for PNI studies, including the utilizations of mouse models and vaccine models. In addition, this book focuses on translational medicine by applying PNI methods in clinical practice. As part of the Methods in Molecular Biology series, the chapters seek to provide a practical, state-of-the-art, and holistic view for the translation of PNI into better preventive and personalized medical practice. Authoritative and hands-on, Psychoneuroimmunology: Methods and Protocols, Second Edition will be of use to biomedical students and professionals at all levels who are interested in integrative studies in psychology, psychiatry, neuroscience, immunology, molecular biology, genetics, bioengineering, physiology, pathology, microbiology, systems biology, and clinical medicine.

The orchid family is one of the largest families of flowering plants known for their beauty and economic importance. This work provides information in key areas of research that are important to both scientists and commercial growers alike. The main purposes of this book are to provide key practical areas of research, such as, germination, micropropagation, traditional and current techniques related to plant improvement; document methods that ensure survival of plants from laboratories to greenhouses; promote communication between scientists and growers, so that their combined expertise on these areas will lead to the successful growth of orchids in their natural habitats or commercial greenhouses. This book can serve as reference for laymen with an interest in orchid growing.This book is divided into 5 parts. The first part emphasizes propagation methods using seeds and related techniques that are important to plant conservation and improvement. Successes in asymbiotic and symbiotic seed germination are keys to orchid conservation and their propagation.  The second part summarizes micropropagation methods, common media, and newer methods of micropropagation such as the bioreactor culture procedures. The third part focuses on techniques related to the manipulation of explants in an in vitro environment. The fourth part covers cell biological methods and transformation techniques. Since the successes in a laboratory setting do not guarantee plant survival and propagation in greenhouses and in the natural environment, it discusses greenhouse propagation techniques that are essential to the survival of plants generated from a laboratory setting. The fifth part showcases recent successes on orchid propagation by documenting sample publications and how to present orchids in an artistic fashion for one’s enjoyment.

Essential in biological functions like cell signaling and, when disturbed, a likely cause of disease, lipids have proven to be a vital force in cell biology. In Lipidomics: Methods and Protocols, an international panel of experts present a wide variety of reviewed as well as unpublished data on isolation techniques, structural analysis, lipid rafts, lipid trafficking and profiling, biomarkers, lipid peroxidation, biostatistics applied to lipids, software tools, and bioinformatics. These studies range from simple systems, such as in yeast, to complex biological models. Samples used in these studies include: mitochondria, microsomes, endosomal membranes, RBS and plasma membranes, vesicles, caveolae, liposomes, macrophages, meibum, brain, microglia, retina, adipose tissue, keratinocytes, pulmonary surfactant, and hepatic cultures. Volume 1 focuses on shotgun and global lipidomics, analytical approaches, and lipid maps. Written in the highly successful Methods in Molecular Biology™ series format, the chapters include useful introductions to their respective topics, lists of the necessary equipment and materials, step-by-step, readily reproducible laboratory protocols, and authoritative notes on troubleshooting and avoiding common pitfalls. Comprehensive and cutting-edge, Lipidomics: Methods and Protocols will support researchers who wish to increase their utilization of lipidomics, which will certainly lead to more powerful technology, improved diagnostic-prognostic capabilities, and the identification of new valuable products.

Homologous recombination is important in various aspects of DNA metabolism, including damage repair, replication, telomere maintenance, and meiosis, and yeast genetics has successfully provided a framework for the mechanism of homologous recombination. Divided into four convenient sections, DNA Recombination: Methods and Protocols covers recent techniques that best utilize the advantages of the yeast system, prescribing to the belief that yeast will keep serving as a great model organism to study homologous recombination. Chapters have also been included for such exceptions as the group of genes involved in recombination that are found solely in higher eukaryotes, such as BRCA2. And looking forward, a necessary step in the direction of understanding the homologous recombination process is to isolate the machine and let it work in a test tube.  Understanding the design by studying the appearance and behavior of the machinery as a single molecule will be an important milestone toward understanding the mechanism of action of the machinery. Techniques covering these topics have also been included.  Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and easily accessible, DNA Recombination: Methods and Protocols serves as an ideal guide to scientists of all backgrounds with its well-honed methodologies and strives to bring the reader to the next level of understanding regarding this vital subject.

This volume presents the latest protocols for both laboratory and bioinformatics based analyses in the field of marine genomics. The chapters presented in the book cover a wide range of topics, including the sampling and genomics of bacterial communities, DNA extraction in marine organisms, high-throughput sequencing of whole mitochondrial genomes, phylogenomics, SNP discovery, SNP-arrays for species identification, digital PCR-based quantification methods, environment DNA for invasive species surveillance and monitoring, microarrays for the detection of waterborne pathogens, DNA barcoding of marine biodiversity, metabarcoding protocols for marine eukaryotes, analytical protocols for the visualization of eukaryotic diversity, and applications for genomic data to benthic indices for environmental monitoring. Written in the highly successful Methods in Molecular Biology series format, chapters include introduction 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.Cutting-edge and thorough, Marine Genomics: Methods and Protocols is a valuable resource for researchers, students, and policy makers in the field of marine biology.

This volume presents the latest protocols for both laboratory and bioinformatics based analyses in the field of marine genomics. The chapters presented in the book cover a wide range of topics, including the sampling and genomics of bacterial communities, DNA extraction in marine organisms, high-throughput sequencing of whole mitochondrial genomes, phylogenomics, SNP discovery, SNP-arrays for species identification, digital PCR-based quantification methods, environment DNA for invasive species surveillance and monitoring, microarrays for the detection of waterborne pathogens, DNA barcoding of marine biodiversity, metabarcoding protocols for marine eukaryotes, analytical protocols for the visualization of eukaryotic diversity, and applications for genomic data to benthic indices for environmental monitoring. Written in the highly successful Methods in Molecular Biology series format, chapters include introduction 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.Cutting-edge and thorough, Marine Genomics: Methods and Protocols is a valuable resource for researchers, students, and policy makers in the field of marine biology.

Peptide synthesis has emerged as one of the most powerful tools in biochemical, pharmacological, immunological, and biophysical la- ratories. Recent improvements include general solid-phase method- ogy, new protecting groups, and automated equipment. These advances have allowed the facile synthesis of increasingly more complex p- tides. Many of these new and improved methods for the synthesis of peptides and peptide-related substances have been reported in various publications, but never compiled in a convenient handbook. Like other volumes in this series, Peptide Synthesis Protocols concentrates on the practical aspects of these procedures, providing the researcher with detailed descriptions and helpful tips about potential problems. This volume is not intended to serve as a basic guide to standard Merrifie- type solid-phase strategy, but rather to provide the researcher with some of the most recent applications in the field of peptide science. A c- panion volume, Peptide Analysis Protocols, will detail methodology for the charaterization of new synthetic peptides. Development of new methods and applications has continued actively even as this volume was in preparation. Owing to the number of contributors to this volume, it was necessary to establish a cutoff for publication purposes. We feel that all of the protocols presented are timely and up-to-date. Several promising new strategies, such as allyloxycarbonyl-based syntheses, were being developed at the time this volume was in the editing stages and will be included in future editions.