Quantitative Real-Time PCR: Methods and Protocols focuses on different applications of qPCR ranging from microbiological detections (both viral and bacterial) to pathological applications. Several chapters deal with quality issues which regard the quality of starting material, the knowledge of the minimal information required to both perform an assay and to set the experimental plan, while the others focus on translational medicine applications that are ordered following an approximate logical order of their medical application. The last part of the book gives you an idea of an emerging digital PCR technique that is a unique qPCR approach for measuring nucleic acid, particularly suited for low level detection and to develop non-invasive diagnosis. Written for the Methods in Molecular Biology series, most chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, laboratory protocols and tips on troubleshooting and avoiding known pitfalls. Practical and authoritative, Quantitative Real-Time PCR: Methods and Protocols aims to aid researchers seeking to devise new qPCR-based approaches related to his or her area of investigation.

This volume provides techniques on recent developments that use RNA scaffolds as molecular tools. RNA Scaffolds: Methods and Protocols guides readers through methods on various domains as molecular biology, nanotechnologies, and structural biology. Written for the Methods in Molecular Biology series, 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.Practical and authoritative, RNA Scaffolds: Methods and Protocols will serve as an invaluable reference for those interested in further study into this fascinating field.

Every year there are new and exciting developments in assisted human reproduction, but how much do we really know about the underlying causes of infertility? This volume explores recent progress in the understanding of the genetics of spermatogenesis and male infertility. Topics include fundamental advances and current problems in the development and function of the testis, an outline of clinical findings in male infertility and an overview of the role of the Y chromosome in male fertility. Comprehensive critiques of posttranscriptional control during spermatogenesis, mammalian meiotic sterility, and comparative genetics of human spermatogenesis from the perspective of yeast, "Drosophila" and mice provide a global overview of the field.

This book presents some of the most recent, novel and fascinating examples of transcriptional and posttranscriptional control of gene expression in plants and, where appropriate, provides comparison to notable examples of animal gene regulation.

The Genetical Theory of Natural Selection by R.A. Fisher (1930) dictated that sexual dimorphisms may depend upon a single medelian factor. This could be true for some species but his suggestion could not take off the ground as gender in Drosophila is determined by the number of X chromosomes. Technical advances in molecular biology have revived the initial thinking of Fisher and dictate that TDF or SRY genes in humans or Tdy in mice are sex determining genes. The fortuitous findings of XX males and XY female, which are generally termed sex reversal phenomenon, are quite bewildering traits that have caused much amazement concerning the pairing mechanism(s) of the pseudoautosomal regions of human X and Y chromosomes at meiosis. These findings have opened new avenues to explore further the genetic basis of sex determination at the single gene level.
The aim of the fourth volume, titled Genetics of Sex Determination is to reflect on the latest advances and future investigative directions, encompassing 10 chapters. Commissioned several distinguished scientists, all pre-eminent authorities in each field to shed their thoughts concisely but epitomise their chapters with an extended bibliography. Obviously, during the past 60 years, the metoric advances are voluminous and to cover every account of genes, chromosomes, and sex in a single volume format would be a herculean task. Therefore, a few specific topics are chosen, which may be of great interest to scientists and clinicians. The seasoned scientists who love to inquire about the role of genes in sex determination should find the original work of these notable contributors very enlightening. This volume is intended for advanced students who want to keep abreast as well as for those who indulge in the search for genes of sex determination.

A one-stop resource that provides the most frequently needed information on the human genome What are the genes that manke-up the human genome, what do the genes do when they are acting properly, and what happens when these genes are damaged? Designed for today's reader who demands quick answers to a wide range of questions, The Human Genome Sourcebook is intended to offer the non-specialist an accessible but detailed guide to the genome. The information it provides is given context: namely, the basic scientific principles of genome research, the new knowledge unearthed or created by this research, and the social and ethical implications of this knowledge. The Human Genome Sourcebook is organized in several sections to simplify the location of pertinent information: an extensive section that comprises an in-depth catalogue of human genes listed according to the roles they play in life; a chapter that relates genetic diseases to the specific genes that cause the disease; a detailed glossary giving readers a deeper understanding of genetic terms and concepts; an overview or roadmap of the physical layout of the genome sections. provides both information as well as the tools necessary to access that information.. Provides in-depth information on the relationships between our genes and all aspects of our daily lives. Addresses health issues that are related to genetic abnormalities

Although they comprise one of the three fundamental branches of life, it was only the last decade that Archaea were formally recognized as a group alongside Eukaryotes and Bacteria. Bacteria-like in that they are single celled organisms that lack a nucleus and intracellular organelles, the Arachaea also share a large gene set typical of eukaryotes, for making and repairing DNA, RNA and protien. More surprisingly, they only inhabit environments typical of the extremes of early earth--hot springs, thermal ocean vents, saline lake, or oxygen deficient sediments. A breakpoint on the common evolutionary path, it is evident that the Archaea diverged early in the history of life, establishing thier importance in evolutionary sciences. "Archaea: Ancient Microbes, Extreme Environments, and the Origin of Life" tells this evolving story, furthering our understanding of the microbe commonalities, and providing for evolutionary justification in the use of archaea as mechanistic model systems.
Key Features
* Provides a unique and current summary of common subcellular mechanisms in archaea and eukaryotes
* Emphasizes the use of genomics to provide a biological context for understanding archaea
* Contrasts evolutionary studies on the fossil record with those on molecular phylogeny
* Includes extensive tables, graphs, images, drawings and other illustrations
* Simplifies the interdisciplinary challenge necessary to understand the significance of archaea

Genetic linkage maps are an increasingly important tool in both fundamental and applied research, enabling the study and deployment of genes that determine important biological traits. This concise introduction to genetic mapping in species with disomic inheritance enables life science graduate students and researchers to use mapping software to produce more reliable results. After a brief refresher on meiosis and genetic recombination, the steps in the map construction procedure are described, with explanations of the computations involved. The emphasis throughout is on the practical application of the methods described; detailed mathematical formulae are avoided and exercises are included to help readers consolidate their understanding. A chapter on recognising and solving problems provides valuable guidance for dealing with real-life situations. An extensive chapter dedicated to the more complex situation of outbreeding species offers a unique insight into the approach required for many economically important and model species, both plants and animals.

In Human Cloning a panel of distinguished philosophers, medical ethicists, religious thinkers, and social critics tackle the thorny problems raised by the now real possibility of human cloning. In their wide ranging reviews, the distinguished contributors critically examine the major arguments for and against human cloning, probe the implications of such a procedure for society, and critically evaluate the "Report and Recommendations of the National Bioethics Advisory Commission." The debate includes both religious and secular arguments, as well as an outline of the history of the cloning debate and a discussion of human cloning's impact on our sense of self and our beliefs about the meaning of life.

This compendium presents some of the major applications of neutron scattering techniques to problems in biology. It is a record of the papers presented at the Neutrons in Biology Conference, the third in an occasional series held to highlight progress in the field and to provide a focus for future direction. The strength ofthe neutron scattering technique remains principally in the manipula tion of scattering density through hydrogen and deuterium atoms. The development ofad vanced detectors, innovative instrument and beamline components, and sophisticated data acquisition systems through the 1970s and early 1980s provided a sound foundation for the technique. With continued development, some of the exotic and expensive equipment has become affordable by the medium-sized facilities, thereby broadening the user base considerably. Despite problems with the major neutron sources in the late 1980s and early 1990s, some spectacular results have been achieved. Whilst the high and medium flux beam reac tors will continue to make a major impact in the field, the results from the first experi ments, and the planned developments on spallation neutron sources, clearly indicate that the technique has enormous potential.

Cells in the developing embryo depend on signals from the extracellular environment to help guide their differentiation. An important mediator in this process is the extracellular matrix -- secreted macromolecules that interact to form large protein networks outside the cell. During development, the extracellular matrix serves to separate adjacent cell groups, participates in establishing morphogenic gradients, and, through its ability to interact directly will cell-surface receptors, provides developmental clocks and positional information. This volume discusses how the extracellular matrix influences fundamental developmental processes and how model systems can be used to elucidate ECM function. The topics addressed range from how ECM influences early development as well as repair processes in the adult that recapitulate developmental pathways. The series Biology of Extracellular Matrix is published in collaboration with the American Society for Matrix Biology.

A compendium of readily reproducible and novel methods to manipulate DNA viruses and characterize their varied biological properties. The authors emphasize techniques for viral detection and genetics, but also include methods for structure determination, gene expression, replication, pathogenesis, complex cellular models, recombinant genetics, and computational/systems approaches. Wide-ranging and highly practical, DNA Viruses: Methods and Protocols will stimulate new directions in virology research with its novel strategies for engineering viral vectors in gene therapy, and its advanced approaches for detecting viruses in human disease.

"Biomimetics and Stem Cells: Methods and Protocols" collects a series of approaches to demonstrate the role and value of biomimetics for the better understanding of stem cell behavior and the acceleration of their application in regenerative medicine. Recent advances in tissue engineering are enabling scientists to instruct stem cells toward differentiating into the right phenotypes, in the right place and at the right time. Given these advances, biomimetic environments are being designed to recapitulate, in vitro, the combinations of factors known to guide tissue development and regeneration in vivo and thereby help unlock the full potential of the stem cells. 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.Practical and essential, "Biomimetics and Stem Cells: Methods and Protocols" focuses on the use of biomimetic systems for stem cells in order to aid in moving this vital field of study forward."

Few would dispute the truth of the statement People are Different', but there is much controversy over why. This book authoritatively explains the methods used to understand human variation, and extends them far beyond the primary nature or nurture' question. After chapters on basic statistics, biometrical genetics, matrix algebra and path analysis, there is a state-of-the-art account of how to fit genetic models using the LISREL package. The authors explain not only the assumptions of the twin method, but how to test them. The elementary model is expanded to cover sex limitation, sibling interaction, multivariate and longitudinal data, observer ratings, and twin-family studies. Throughout, the methods are illustrated by applications to diverse areas such as obesity, major depression, alcohol comsumption, delinquency, allergies, and common fears.

Splicing of primary RNA transcript is a quasi-systematic step of gene expression in higher organisms. This is the first book to highlight the medical implications, i.e. diseases, caused by alternative splicing. Alternative splicing not only vastly increases protein diversity but also offers numerous opportunities for aberrant splicing events with pathological consequences. The book also outlines possible targets for therapy.

The last 15 years in development of biology were marked with accumulation of unprecedentedly huge arrays of experimental data. The information was amassed with exclusively high rates due to the advent of highly efficient experimental technologies that provided for high throughput genomic sequencing; of functional genomics technologies allowing investigation of expression dynamics of large groups of genes using expression DNA chips; of proteomics methods giving the possibility to analyze protein compositions of cells, tissues, and organs, assess the dynamics of the cell proteome, and reconstruct the networks of protein-protein interactions; and of metabolomics, in particular, high resolution mass spectrometry study of cell metabolites, and distribution of metabolic fluxes in the cells with a concurrent investigation of the dynamics of thousands metabolites in an individual cell. Analysis, comprehension, and use of the tremendous volumes of experimental data reflecting the intricate processes underlying the functioning of molecular genetic systems are unfeasible in principle without the systems approach and involvement of the state-of-the-art information and computer technologies and efficient mathematical methods for data analysis and simulation of biological systems and processes. The need in solving these problems initiated the birth of a new science- postgenomic bioinformatics or systems biology in silico.

The in situ hybridization and PCR technologies are now well-established molecular techniques for studying chromosomal aneuploidy and rearran- ments, gene localization and expression, and genomic organization. Over the last decade, we have seen increasing applications in these fields. By combining the high sensitivity of the PCR reaction and the cytological localization of target sequences, both PRINS and in situ PCR techniques have provided highly powerful complements to FISH for in situ cellular and molecular investigations. Both these approaches have several advantages in terms of sensitivity and specificity, owing to the use of primers and to the fast kinetics of annealing and elongation reactions in situ. In the first edition of PRINS and In Situ PCR Protocols edited by John R. Gosden, experts in the field presented in detail a variety of applications of PRINS and in situ PCR techniques, in a wide range of clinical conditions. Since the publication of this successful reference book, there have been s- nificant improvements in in situ detection techniques. This completely revised and updated second edition presents a compreh- sive selection of new procedures developed in the field of PRINS and in situ PCR technologies. The book has two sections. Part I, Basic Methodology, contains chapters that provide useful protocols for many variations of PRINS and in situ PCR, including a new fast multicolor PRINS method, and protocols for PRINS detection of unique sequences in situ.

RNA Biochemistry and Biotechnology describes various aspects of nucleic acid and protein structure, mainly RNA structure and proteins, interacting with specific RNA species. Papers deal with DNA protein interactions, telomerase, aminoacyl-tRNA synthetases, elongation factor Tu, DNA repair, RNA structure, NMR technology, RNA aptamer interaction of biological macromolecules with metal ions. Two papers deal with theoretical aspects of RNA structure production and computer modelling. Many papers describe the possibility of commercial application of RNA biotechnology. One article discusses the impact of direct democracy on basic science supporting biotechnology. Readership: Advanced graduate students, Ph.D. students and young scientists as well as specialists in the field.

nd During June 13 -June 23 1996, the 2 EL. B. A. Foundation course on Genome, a NATO Advanced Study Institute, was held at Marcian Marina, Isle of Elba, Italy, - sponsored by the North Atlantic Treaty Organization and the EL. B. A. Fundation. The subject of the course was "Genome Structure and Function" with participants selected worldwire from 15 afferent countries. The purpose of the course and of the resulting book is the study of DNA structure (from the primary to the quintemary) and gene expression in the control of cell function and cell cycle progression; the topics were presented by top experts, covering both structural (cbwn to the atomic resolution) and functional (cbwn to gene level) aspects. The topics were presented by top experts and scientists active in the field, with the goal to give an insight into modm problems of genome study and recent ochievements in related fielm of molecular and cell biology, genetic engineering, biochemistry and biophysics, oncology and biotechnology. This resulting book is intenred to give a broad perspecti ve of the current stand of these fields. The major emphasis is towarm a reep unrerstanang of DNA structure and function in intetphase and metaphase chromosomes, originating by the parallel biophysical (namely NMR X-Ray and neutron scattering, spectropolarimetry, image analysis, calorimetry) and biochemical study conwcted on a wire range of cell systems placing the emphasis on either the higher orrer DNA structure or gene structure and function.

This book updates and expands on various aspects of the vasculature's microenvironment and how these regulate differentiation and assembly. Discussed in this new edition are efforts to capitalize on combing engineering techniques, to study and manipulate various biophysical cues, including: endothelial cell- pericyte interactions (Davis), mechanical forces to regulate vascularization in three-dimensional constructs (Levenberg), how matrix properties and oxygen tension regulate vascular fate and assembly (Gerecht), biophysical cues in relation to vascular aging (Ferreira), 3D printing of complex vascularized tissue (Hibino), the harnessing of biophysical cues for therapeutic vasculature interfacing with the damaged brain (Segura) and finally, the infarcted heart (Grayson). This second edition of Biophysical Regulation of Vascular Differentiation and Assembly provides an interdisciplinary view of vasculature regulation thru various biophysical cues and presents recent advances in measuring and controlling such parameters. This book will be of interest to biologists, biophysicists and engineers who work with vascular differentiation and assembly.

This volume brings together the disciplines of plant and animal genome research, and serves as an opportunity for scientists from both fields to compare results, problems and prospects.

Legionellosis is a disease of significant medical and public interest. Legionella is commonly found in aquatic habitats where its ability to survive and to multiply within different protozoa equips the bacterium to be transmissible and pathogenic to humans. In addition, Legionella has become a favored model system to analyze the mechanisms of bacterial survival, acquisition of nutrients, and intracellular replication. Following the recent publication of the genome sequences of four L. pneumophila strains, it is now feasible to investigate the whole genome in silico, the transcriptome via micro arrays, and the proteome by two-dimensional gel electrophoresis. Research in the fields of clinical features, diagnosis, treatment, and epidemiology continues to generate new data. The topics covered by this volume range from the history of the identification of Legionella and clinical disease treatment, to the microbe's gene expression and secretion systems, as well as its strategies for intracellular multiplication and nutrient acquisition. The main focus of the book is the current state of many of the most critical features of Legionella. Internationally renowned authors have contributed chapters describing and discussing the latest research findings with an emphasis on molecular aspects. The editors and authors have produced an excellent book that will be an extremely useful reference source. This comprehensive publication is aimed at readers with teaching or research interests in microbiology, genetics, genomics, infectious diseases, or clinical research.

The intersection of race, ethnicity and genomics has recently been a focus of debate and concern. The key areas of debate are pharmacogenomics and, to a lesser extent, racial profiling in the criminal justice system. The former poses the question as to whether certain "races" are genetically predisposed towards given diseases and whether they metabolize drugs differently; with the latter debating whether DNA analyses accurately identify the "race" of an individual. This book takes a different approach, while acknowledging the importance of these debates and their role in shaping what the issues are perceived to be in thinking about the intersection of race, ethnicity and genomics. We are interested in exploring the interconnections between race, ethnicity and nation and kinship, always bearing in mind that kinship, as a domain of human experience and a field of social study, has been reshaped by the genomic and biotechnological revolution. Peter Wade is Professor of Social Anthropology at the University of Manchester. His publications include Blackness and Race Mixture (1993), Race and Ethnicity in Latin America (1997), Music, Race and Nation: Musica Tropical in Colombia (2000), Race, Nature and Culture: An Anthropological Perspective (2002). His current research focuses on issues of racial identity, embodiment and new genetic and information technologies.

There can be no doubt that some ofthe most spectacular advances made in science over the past few decades have been in the isolation, analysis, and manipulation of nucleicacids. Thishas ledtoamuchgreaterunderstandingofmechanismsandprocesses across many fields of bioscience, such as biochemistry, microbiology, physiology, pharmacology, and the medical sciences to name a few. It has also led to the growth of the biotechnology industry, which seeks to develop and commercialize many ofthese important processes and methods. Much ofthis has come about because ofthe devel opment of numerous molecular biology and genetic manipulation techniques. The discovery of restriction enzymes and the development of cloning vectors in the early 1970sopenedthedoortowaysofisolatingandmanipulatingnucleic acidsthathadnever been thought possible. Gene probe labeling and hybridization were developed and refined toprovidepowerfulmethodsofanalysis. These-togetherwiththedevelopment of DNA sequencing methods, protein engineering techniques, and PeR-have all continued to contribute substantially to the understandingofbiological processes at the molecular level. Theprotocols for these importantmethods are the focus ofThe Nucleic AcidProtocols Handbook, whose aim is to provide a comprehensive set oftechniques in onevolume thatwill enable the isolation, analysis, and manipulationofnucleic acids to be readily undertaken. The NucleicAcidProtocols Handbook is divided into 10 parts; within each there are approximately 10chapters. The first fourpartsfollow oneanotherlogically: nucleic acid extraction (Part I), basic separation and analysisofDNA (II), through probe design and labeling (III), and RNA analysis techniques (IV). The following three sections deal with gene libraryconstruction andscreening(V), DNA sequencing (VI), andthe polymerase chain reaction (VII)."

The potential now exists in many experimental systems to transfer a cloned, modified gene back into the genome of the host organism. In the ideal situation, the cloned gene is returned to its homologous location in the genome and becomes inserted at the target locus. This process is a controlled means for the repair of DNA damage and ensures accurate chromosome disjunction during meiosis. The paradigm for thinking about the mechanism of this p- cess has emerged primarily from two sources: (1) The principles of reaction mechanics have come from detailed biochemical analyses of the RecA protein purified from Escherichia coli; and (2) the principles of information transfer have been derived from genetic studies carried out in bacteriophage and fungi. A compelling picture of the process of homologous pairing and DNA strand exchange has been influential in directing investigators interested in gene t- geting experiments. The ability to find and pair homologous DNA molecules enables ac- rate gene targeting and is the central phenomenon underlying genetic recombi- tion. Biochemically, the overall process can be thought of as a series of steps in a reaction pathway whereby DNA molecules are brought into homologous register, the four-stranded Holliday structure intermediate is formed, hete- duplex DNA is extended, and DNA strands are exchanged. Not much is known about the biochemical pathway leading to homologous recombination in euka- otes.