Real-time PCR (RT-PCR) technology is highly flexible and many alternative instruments and fluorescent probe systems have been developed recently. The decreased hands-on time, increased reliability, and improved quantitative accuracy of RT-PCR methods have contributed to the adoption of RT-PCR for a wide range of new applications. This essential manual presents a comprehensive guide to the most up-to-date technologies and applications, as well as providing an overview of the theory of this increasingly important technique. Renowned experts in the field describe and discuss the latest PCR platforms, fluorescent chemistries, validation software, data analysis, and internal and external controls. This timely and authoritative volume also discusses a wide range of RT-PCR applications including clinical diagnostics, biodefense, RNA expression studies, validation of array data, mutation detection, food authenticity and legislation, NASBA, molecular halotyping, and much more. Real-Time PCR: Current Technology and Applications will be an essential book for all laboratories using PCR.

Population and evolutionary genetics have been quickly developing ?elds of biological research over the past decades. This book compiles our current understanding of genetic processes in natural populations. In addition, the book provides the author's original ideas and concepts based on the data obtained by himself and his close coworkers. The author introduces his pioneering concept of population genetic stability, and much of thebook is concerned with the factors and conditions of such stability. Why does genetic stability matter so much? Altukhov argues that the sustainable use of natural resources, including genetic resources of popu- tions, critically depends on the maintenance of their stability. The preser- tion of well-adapted genetic characteristics from one generation to the next is essential for this stability. Traditionally, population genetics has been - cusedonevolution andthe role of evolutionary factorsinshapinggenetic structures of populations. While the idea of a population as a dynamic unit of evolution has been widely accepted, the signi?cance of genetic stability and its implications for the long-term survival of populations and species have not been fully appreciated.

This book explores Dental Stem Cell (DSC) biology, from a review of basic concepts for cell culture, to isolation, self-renewal, multipotency and differentiation, regulation by molecular medicine, and prospective research areas for regenerative medicine. The first seven chapters delve into basic DSC properties, vital signaling pathways involved in differentiation, pluripotency, iPS cell development from DSCs, and genetic engineering approaches of DSCs in accordance with the current literature. A comprehensive review of possible clinical applications and in vitro/in vivo studies follows, illustrating the future of DSC research for in the tissue engineering field. The text also discusses the political, ethical, social, and legal ramifications of the use of dental stem cells. Expertly authored and drawing from a multitude of international perspectives, Dental Stem Cells is an invaluable addition to Springer's Stem Cell Biology and Regenerative Medicine series. It is essential reading for advanced graduate students, basic researchers, and clinical investigators in the fields of stem cell therapy, biological sciences of dentistry, and regenerative medicine.

The second edition of this book adds eight new contributors to reflect a modern cutting edge approach to genomics. It contains the newest research results on genomic analysis and modeling using state-of-the-art methods from engineering, statistics, and genomics. These tools and models are then applied to real biological and clinical problems. The book 's original seventeen chapters are also updated to provide new initiatives and directions.

This is the first book to examine organelle proteomics in depth.

It begins by introducing the different analytical strategies developed and successfully utilized to study organelle proteomes, and detailing the use of multidimensional liquid chromatography coupled to tandem mass spectrometry for peptide sample analysis. Detailed protocols are provided and a section is devoted to methods enabling a global estimate of the reliability of the protein list assigned to an organelle.

The efficiency of delivering DNA into mammalian cells has increased t- mendously since DEAE dextran was first shown to be capable of enhancing transfer of RNA into mammalian cells in culture. Not only have other chemical methods been developed and refined, but also very efficient physical and viral delivery methods have been established. The technique of introducing DNA into cells has developed from transfecting tissue culture cells to delivering DNA to specific cell types and organs in vivo. Moreover, two important areas of biology-assessment of gene function and gene therapy-require succe- ful DNA delivery to cells, driving the practical need to increase the efficiency and efficacy of gene transfer both in vitro and in vivo. TM These two volumes of the Methods in Molecular Biology series, Gene Del- ery to Mammalian Cells, are designed as a compendium of those techniques that have proven most useful in the expanding field of gene transfer in mammalian cells. It is intended that these volumes will provide a thorough background on chemical, physical, and viral methods of gene delivery, a synopsis of the myriad techniques currently available to introduce genes into mammalian cells, as well as a practical guide on how to accomplish this. It is my expectation that it will be useful to the novice in the field as well as to the scientist with expertise in gene delivery.

Over the last decades, scientists have been intrigued by the fascinating organisms that inhabit extreme environments. These organisms, known as extremophiles, thrive in habitats which for other terrestrial life-forms are intolerably hostile or even lethal. Based on such technological advances, the study of extremophiles has provided, over the last few years, ground-breaking discoveries that challenge the paradigms of modern biology. In the new bioeconomy, fungi in general, play a very important role in addressing major global challenges, being instrumental for improved resource efficiency, making renewable substitutes for products from fossil resources, upgrading waste streams to valuable food and feed ingredients, counteracting life-style diseases and antibiotic resistance through strengthening the gut biota, making crop plants more robust to survive climate change conditions, and functioning as host organisms for production of new biological drugs. This range of new uses of fungi all stand on the shoulders of the efforts of mycologists over generations. The book is organized in five parts: (I) Biodiversity, Ecology, Genetics and Physiology of Extremophilic Fungi, (II) Biosynthesis of Novel Biomolecules and Extremozymes (III) Bioenergy and Biofuel synthesis, and (IV) Wastewater and biosolids treatment, and (V) Bioremediation.

In "Animal Models for Stem Cell Therapy: Methods and Protocols, "expert researchers in the field detail disease models of hepatic, cardiovascular, neurological diseases, connective and contractile tissue. Chapters focus on a wide range of diseases and application of different kinds of stem cells and reprogrammed tissue cells (iPS).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 key tips on troubleshooting and avoiding known pitfalls.

Authoritative and practical, "Animal Models for Stem Cell Therapy: Methods and Protocols, "covers interest of basic scientists and clinicians to assess the biological as well as the therapeutic potential of stem cell therapy."

Based on the international workshop GPCRs: From Deorphanisation to Lead Structure Identification, held in Berlin in May 2006, the book highlights the following topics: Structure of GPCRs, Design of GPCR Ligands, GPCR Signalling, Deorphanization and Assay Development. All chapters are written by leading experts in the field, discussing the most recent state of the art. They give insight into the approaches taken by industry and academia to address GPCRs and depict how mature this target class-oriented research has become in the last decade. The book reflects the actual trends in the fast-emerging field of GPCR research in academia and industry.

Stem Cell therapy for lysosomal diseases (LSDs) is developing rapidly. This volume discusses the history, current practice and future perspectives of stem cells in inborn errors of metabolism (IEM) and provides an international perspective on progress, limitations, and future directions (e.g. gene therapy, iPS, ES) in the field. Beginning with an overview of these diseases, the book covers the breadth of this topic from treatment options, bone marrow transplantation, and alternative treatment options, through long-term outcomes and future perspectives.

This volume summarizes recent advances in research on mesenchymal cell populations in the bone marrow. It explores how mesenchymal cells create niches for immune cells in extramedullary organs and it discusses new concepts of lympho-hematopoietic microenvironments. Readers are introduced to the fundamentals of hematopoietic stem cells (HSCs) differentiation to all types of blood cells, including immune cells, in the bone marrow. The book highlights how this process is supported and regulated by the individual microenvironments of stem cells, termed niches. The identity of HSC niches has been subject to longstanding debates. Recent studies identified the population of mesenchymal stem cells as the major cellular component of niches, for hematopoietic stem and progenitor cells (HSPCs) and their candidate developmental origin. Furthermore, candidate cellular niches for immune cells in lymph nodes and adipose and connective tissues were identified. The authors of this volume focus on shared features between those and HSPC niche cells in the bone marrow. Covering latest research results, this book serves as fascinating read for researchers and clinicians in hematology and immunology.

The book covers various biotechnological research efforts and their applications in fisheries and aquaculture, especially in the area of fish breeding, health management, nutrition and culture. Application of the recent biotechnological tools, like Transcriptomics, Transgenesis, Nanotechnology, Metabolomics, RNAi and CRISPRi Technologies in the field of fisheries research are included in the book. Topics like conservation genetics for management of fishery resources are also covered in the book. It aims at addressing the growing need of the biotechnology in advancing the cause of aquaculture with a view to provide food and nutritional security to the world. This book will be of immense use to teachers, researchers, academicians, development officials and policymakers, involved in R&D of fisheries and aquaculture sectors. Also, the book serves as an additional reading material for undergraduate and graduate students of fisheries, marine sciences, ecology, aquaculture, and environmental sciences. The research in aquaculture biotechnology is likely to have significant impact on aquaculture and fisheries by way of supporting nutritional food security to the growing population.

This book contains a comprehensive collection of experimental and computational strategies and techniques for microbial genome-scale essentiality studies, developed and presented by the leading groups in the field. It contains detailed description of the procedures, discussion of potential difficulties and failures. All protocols follow the successful Methods in Molecular Biology(TM) series format.

Artificial riboswitches and other ligand-responsive gene regulators make it possible to switch protein synthesis ON or OFF with arbitrary ligand molecules. Artificial Riboswitches: Methods and Protocols focuses on the state-of-the-art methods developed in recent years for creating artificial riboswitches, therefore this volume could be regarded as a collection of recipes for the gene circuit elements in synthetic biology and metabolic engineering. Chapters cover topics such as screening or rational design methods for obtaining artificial riboswitches that function in either bacterial or eukaryotic translational systems, protocols for evaluating the activities of the resultant riboswitches, as well as protocols for construction of ligand-dependent, trans-acting gene regulators. 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, Artificial Riboswitches: Methods and Protocols seeks to serve not only bioengineers who aim to reprogram cell behaviors and molecular biologists who leverage these regulators for genetic studies, but to all researchers interested in this fascinating field.

Genetic, hormonal, neurological, and other biological factors need to be taken into account to fully understand sexual orientation. This work represents the latest research and theory on causes of variation in sexual orientation. It looks at sexual orientation as a cross-species phenomenon with numerous determining factors.

This work is a collection of chapters by some of the leading researchers in the scientific study of sexual orientation. The theory that many genetic, hormonal, neurological, and other biological factors need to be taken into account to fully understand sexual orientation is espoused in this book. It presents much of the latest research on the causes of variation in sexual orientation and related phenomena. It views sexual orientation as a cross-species phenomenon with both biological and environmental determinants.

"Directed Evolution Library Creation: Methods and Protocols, Second Edition "presents user-friendly protocols for both proven strategies and cutting-edge approaches for the creation of mutant gene libraries for directed evolution. As well as experimental methods, information on current computational approaches is provided in a user-friendly format that will allow researchers to make informed choices without needing to comprehend the full technical details of each algorithm. Directed evolution has become a fundamental approach for engineering proteins to enhance activity and explore structure-function relationships, and has supported the rapid development of the field of synthetic biology over the last decade. Divided into three convenient sections, topics include point mutagenesis strategies, recombinatorial methods wherein genetic diversity is sourced from multiple parental genes that are combined via either homology-dependent or -independent techniques and a variety of computational methods to guide the design and analysis of mutant libraries. 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, "Directed Evolution Library Creation: Methods and Protocols, Second Edition "will serve as a reliable manual for both novice and experienced protein engineers and synthetic biologists and will enable further technical innovation and the exploitation of directed evolution for a deeper understanding of protein design and function.

Recent stem cell research has revealed that miRNA and RNAi-mediated gene regulation is one of the vital determinates controlling the state of cell differentiation, with the small RNAs serving as key elements involved in regulatory network control of pluripotent cell fate determination. In RNAi and microRNA-Mediated Gene Regulation in Stem Cells: Methods, Protocols, and Applications, expert authors from laboratories across the globe contribute an accessible compendium of up-to-date, proven methods focused on the study of the titular topic. Divided into three sections, the book first gives a brief introduction to RNAi and miRNAs in stem cells, with a focus on the current status of research and future perspectives, then it continues with detailed methods and protocols for RNAi screening, transfection, and the knockdown of specific genes and pathways in several animal species, including humans and mice, concluding with a section on recently developed methods for identification of miRNAs, including a general protocol for preparation and analysis of miRNA libraries for deep sequencing, knock down of a specific gene using miRNA-based shRNA, and miRNA expression analysis using qRT-PCR. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes highlighting tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, RNAi and microRNA-Mediated Gene Regulation in Stem Cells: Methods, Protocols, and Applications serves as a valuable resource for scientists and aspiring graduate students interested in the intersection of RNAi, miRNA, and stem cell molecular biology and the exciting areas of medicine, including regenerative medicine, aging, cancer, and neurological disorders, that can be advanced through this expanding area of research.

Bacteria pathogenic for plants are responsible for devastating losses in agriculture. The use of antibiotics to control such infections is restricted in many countries due to worries over the evolution and transmission of antibiotic resistance. The advent of genome sequencing has enabled a better understanding, at the molecular level, of the strategies and mechanisms of pathogenesis, evolution of resistance to plant defense mechanisms, and the conversion of non-pathogenic into pathogenic bacteria. In this book, internationally acclaimed experts review the most important developments, providing an invaluable up-to-date summary of the molecular biology and genomics of plant pathogenic bacteria. The book opens with two chapters on bacterial evolution, diversity, and taxonomy - topics that have been transformed by molecular biology and genomics analyses. The third chapter delves into the crucially understudied area of pathogen adaptation to the plant apoplast environment. The next seven chapters focus on specific plant pathogens: Agrobacterium, Leifsonia, Pectobacterium, Pseudomonas, Ralstonia, Xanthomonas, and Xylella. The following four chapters review specific, intensively studied areas of research in the plant pathogen field: microbe associated molecular patterns (MAMPs) and innate immunity; use of bacterial virulence factors to suppress plant defense; cyclic di-GMP signalling and the regulation of virulence; and plasmids and the spread of virulence. The final chapter covers the critical area of bioinformatics. With contributions from some of the pioneering bacterial plant pathogen genome sequencers, this book is essential reading for every plant pathogen researcher - from the PhD student to the experienced scientist - as it provides a timely review of the current and most topical areas of research.

This collection reviews developments in DNA profiling across jurisdictions with a focus on scientific and technological developments as well as their political, ethical, and socio-legal aspects. Written by leading scholars in the fields of social studies of forensic science, science and technology studies and socio-legal studies, the book provides state-of-the-art analyses of forensic DNA practices in a diverse range of jurisdictions, new and emerging forensic genetics technologies and issues of legitimacy. The work articulates the various forms of technolegal politics involved in the everyday, standardised and emerging practices of forensic genetics and engages with the most recent scholarly and policy literature. In analyses of empirical cases, and by taking into account the most recent technolegal developments, the book explores what it means to live in a world that is increasingly governed through anticipatory crime control and its related risk management and bio-surveillance mechanisms, which intervene with and produce political and legal subjectivities through human bodies in their DNA. This volume is an invaluable resource for those working in the areas of social studies of forensic science, science and technology studies, socio-legal studies, sociology, anthropology, ethics, law, politics and international relations.

With a particular emphasis on tumor dormancy in breast, lung, prostate, and liver cancers, as well as in melanoma, this first volume of a new Springer series focuses on the interrelationship between biological processes of aging and tumors -- both dormant and quiescent. With detail supplied by numerous international researchers at the forefront of cancer research, the book examines a host of differing aspects of the topic. Featured contributions analyze the role of the quiescent state in regulating hematopoietic and muscle stem cells. They also explore the mediation, by the kinase, in the reversible quiescent state of a subset of ovarian, pancreatic, and colon cancers. The book includes key research on the molecular mechanisms underlying stress-induced cellular senescence, in addition to those governing the accumulation of reactive oxygen species, and the induction of premature senescence. It also provides information on suppressing cellular senescence in the most common, and most aggressive malignant primary brain tumor in humans, glioblastoma multiforme. With comprehensive and cutting-edge information on therapeutic interventions and on the correct diagnosis of relevant neoplasms, and with numerous color illustrations, this is the most up-to-date assessment of current medical knowledge in this crucial area of medical research.

Chromosomes Today volume 14 records the plenary proceedings of the 14th International Chromosome Conference, presenting an overview of the current concerns in plant, animal and human cytogenetics. This volume provides up-to-date information regarding relevant aspects on structure, function and evolution of chromosomes, meiosis, sex chromosomes, and cancer cytogenetics. It contains invited contributions from some of the world's leading experts in the field.

Here is a manual for an environmental scientist who wishes to embrace genomics to answer environmental questions. The volume covers: gene expression profiling, whole genome and chromosome mutation detection, and methods to assay genome diversity and polymorphisms within a particular environment. This book provides a systematic framework for determining environmental impact and ensuring human health and the sustainability of natural populations.

Population genetics occupies a central role in a number of important biological and social undertakings. It is fundamental to our understanding of evolutionary processes, of plant and animal breeding programs, and of various diseases of particular importance to mankind.

This is the first of a planned two-volume work discussing the mathematical aspects of population genetics, with an emphasis on the evolutionary theory. This first volume draws heavily from the author's classic 1979 edition, which appeared originally in Springer's Biomathematics series. It has been revised and expanded to include recent topics which follow naturally from the treatment in the earlier edition, e.g., the theory of molecular population genetics.

This book will appeal to graduate students and researchers in mathematical biology and other mathematically-trained scientists looking to enter the field of population genetics.

Whilst genetic transformation of plants is commonly viewed as a means of bringing about plant improvement, it has not so readily been recognised as a tool for analysing the function of plant genes. This book is unusual in that it focuses on the genetic transformation of a range of plants using a number of different methods. Many plants have been found to be quite difficult to transform, and so various techniques were developed. These techniques include: Agrobacterium suspension drops, electroporation, PEG, "whiskers", and various biolistic methods. A chapter on intellectual and property rights is included.