The polymerase chain reaction (PCR) is a fundamental tool in scientific research and clinical testing. Real-time PCR, combining both amplification and detection in one instrument, is a rapid and accurate method for nucleic acid detection and quantification. Although PCR is a very powerful technique, the results achieved are valid only if the appropriate controls have been employed. In addition, proper optimization of PCR conditions is required for the generation of specific, repeatable, reproducible, and sensitive data. This book discusses the strategies for preparing effective controls and standards for PCR, when they should be employed, and how to interpret the information they provide. It highlights the significance of optimization for efficiency, precision, and sensitivity of PCR methodology and provides essential guidance on how to troubleshoot inefficient reactions. Experts in PCR describe design and optimization techniques, discuss the use of appropriate controls, explain the significance of standard curves, and explore the principles and strategies required for effective troubleshooting. The book highlights the importance of sample preparation and quality, primer design, controlling inhibitors, avoiding amplicon and environmental contamination, optimizing reagent quality and concentration, and modifying the thermal cycling protocol for optimal sensitivity and specificity. In addition, specific chapters discuss the history of PCR, the choice of instrumentation, the applications of PCR in metagenomics, high resolution melting analysis, the MIQE guidelines, and PCR at the microliter scale. The strategies, tips and advice contained in this concise volume will enable the scientist to optimize and effectively troubleshoot a wide range of techniques, including PCR, reverse transcriptase PCR, real-time PCR, and quantitative PCR. It will be an essential book for anyone using PCR technology.

Volume 608 of the series Methods in Enzymology covers key aspects of enzyme discovery, engineering tools and platforms, and examples of applications in the enzymology of synthetic biology.

Microfluidics in Cell Biology Part B: Microfluidics in Single Cells, Volume 147, a new volume in the Methods in Cell Biology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. Unique to this updated volume are three sections on microfluidics in various single cell models, including microfludics in micro-organisms, microfluidics for cell culture and cell sorting of mammalian cells, and microfluidics for cell migration. Specific sections in this latest release include Temperature control and drug delivery for cell division cycle control in fission yeast H2O2 stress response in budding yeast, Antibiotic resistance in bacteria, Metabolism in bacteria, Fluidized beds for bacterial sorting and amplification, Microfluidics for cell culture and cell sorting of mammalian cells, Hydrogel microwells, Immune cells migration in complex environments, Neutrophiles migration in health and disease, Cell guidance by physical cues, Stable gradients in gels of extracellular matrix for cancer cell migration, and more.

Computational Molecular modelling in Structural Biology, Volume 113, the latest release in the Advances in Protein Chemistry and Structural Biology, highlights new advances in the field, with this new volume presenting interesting chapters on charting the Bromodomain BRD4: Towards the Identification of Novel Inhibitors with Molecular Similarity and Receptor Mapping, and Computational Methods to Discover Compounds for the Treatment of Chagas Disease.

An accompanying volume (Volume 6) in this series presents strategies of cellular invasion from the viewpoint of the microbe.
This filed of study is growing rapidly after a somewhat slow start over recent decades. This collection of invited chapters attempts to reflect current research, and brings together cell biologists, microbiologists and immunologists with disparate interests. However, there is a certain unity, even repetition of key themes, hopefully like a symphony rather than a boring catalogue. It will be evident that editorial bias favors intracellular paratism and medically important organisms. The neutrophil is far more than a supporting player to the macrophage, and some attempt is made to remind the reader of some of its unique skills. To retain a manageable size, the emphasis is on relatively early events such as mutual recognition, cell entry, and response, rather than on longterm changes in gene expression by either host cell or pathogen. Viruses are excluded not because of lack of importance but because of somewhat different research approaches, although it is cytogenes, share common strategies in invasion and intercellular spread.

Progress in Molecular Biology and Translational Science, Volume 159, provides the most topical, informative and exciting monographs available on a wide variety of research topics related to prions, viruses, bacteria and eukaryotes. The series includes in-depth knowledge on molecular biological aspects of organismal physiology, along with insights on how this knowledge may be applied to understand and ameliorate human disease. New chapters in this release discuss timely topics, such as Targeting recently deorphanized GPR83 for the treatment of infection, stress, and drug addiction, Arrestin Structure-Function, Arrestins in the Cardiovascular System, Analysis of biased agonism, and more.

Electrophoresis is a straightforward but informative analytical method used in biochemistry, biology and medicine. This book combines a detailed discussion of theory and technical application with an elaborate section on troubleshooting and problem solving in electrophoresis. Therefore the book is an important guide for both students and scientists.

Apoptosis, or programmed cell death, is an adaptive form of cell death that plays a critical role in turnover of mitotic cells and various tissues in the adult, including epithelial cells, fibroblasts and various endocrine cells. Programmed cell death also plays a major role in development in organizing the body plan and molding intricate cellular structures such as nerve cell circuits in the brain. Rapidly progressing research into the molecular and biochemical underpinnings of the programmed cell death process are revealing novel genetic programs and molecular interactions that coordinate a process that results in death and removal of cells without an immune response and in the absence of the adverse effects on neighboring cells.

"Programmed Cell Death, Volume I," critically details the molecular, biochemical and cellular mechanisms of apoptosis. This volume covers programmed cell death in a variety of tissues and organ systems highlighting the interesting families of proteins involved in promoting or preventing apoptosis. These include the caspase and calpain families of proteases, Bcl-2 family members, and inhibitors of apoptosis proteins. Each chapter is written by an internationally recognized expert in a particular aspect of programmed cell death.

This book will provide the reader with a comprehensive understanding of the cascade of events leading from an apoptotic signal, such as trophic factor withdrawal or increased oxidative stress, to cell death. Importantly, this volume also covers signaling mechanisms designed to prevent apoptosis. Such anti-apoptotic signaling cascades involve neurotrophic factors and stress response pathways. "Programmed Cell Death, Volume I," provides the molecular and cellular foundation for http: //www.elsevier.com/locate/isbn/0444507302Programmed Cell Death, Volume II in which the roles of aberrant regulation of apoptosis in human diseases ranging from cancer to Alzheimer's disease are considered.

Retrovirus-Cell Interactions provides an up-to-date review of the interactions between retroviruses and the cells they infect, offering a comprehensive understanding of how retroviruses hijack cellular factors to facilitate virus replication. Drugs targeting viral enzymes have been developed to treat HIV; the next challenge is to inhibit virus-cell interactions as next generation treatment strategies. Organized according to the retrovirus' replication cycle, this book does not focus exclusively on HIV, but rather includes important findings in other retroviral systems, including animal retroviruses, retrotransposons, and endogenous retroelements to allow broad comparisons on important commonalities and differences.

Gangliosides in Health and Disease, Volume 156, presents the latest information on Gangliosides, a class of glycolipids that are found on all vertebrate cell surfaces, and are particularly abundant in the brain. Individual chapters in this new volume cover Gangliosides as Toxin Receptors, Gangliosides in Cancer Cell Signaling, Gangliosides in inflammation and neurodegeneration, Gangliosides as functional galectin receptors, Gangliosides in signal transduction, Gangliosides in brain tumor immunology, and Gangliosides in axon regeneration and stability, amongst other related topics. This book brings together world experts in ganglioside structure and function who have been assembled to contribute to this thorough update of the field.

Epigenetics and Psychiatric Disease, Volume 157, the latest volume in the Progress in Molecular Biology and Translational Science series, includes recent developments on a variety of topics, including the Epigenetic landscapes of the adversity-exposed brain, Chromosomal conformations and epigenomic regulation in schizophrenia, Progress in the epigenetics of depression, the epigenetics of circadian rhythms in imprinted neurodevelopmental disorders, DNA methylation mediating substance abuse, mechanisms and therapeutic opportunities, DNA methylation in animals model of psychosis, Epigenetics of early life stress, Epigenetic drugs for mood disorders, and more.

Methods in Enzymology, Volume 607: Phosphatases, the latest release in this ongoing series, highlights new advances in the field as detailed by an international board of authors. This latest release includes chapters on Empirical Valence Bond Simulations of the Evolution of Enzyme Function, QM/MM Free Energy and Kinetic Isotope Analysis of Phosphoryl Transfer in Enzymes, the Structural, Mechanism and Evolution of Phosphatases, How to Define Rapid Motions in Pumping Pyrophosphatases, The Evolution of K+-Independence in Pyrophosphatases, the Crystallization of Michaelis, Intermediate and Inhibited Complexes in Phosphatases, and an Investigation of Nucleotide Loading and Effector Binding of K-Ras.

International Review of Cell and Molecular Biology, Volume 337 reviews and details current advances in cell and molecular biology. The IRCMB series has a worldwide readership, maintaining a high standard by publishing invited articles on important and timely topics that are authored by prominent cell and molecular biologists. Sections in this new release include the karyosphere (karyosome) and its peculiar structure of the oocyte nucleus, organoids as models of disease, lipid droplets as organelles, the dark side of apoptosis, interconnections between autophagy and secretion, and the regulation and function of intracellular pressure in cell biology.

Bacteria have evolved with extraordinary abilities to regulate aspects of their behavior (such as gene expression) in response to signals in the intracellular and extracellular environment. Key to this are the diverse macromolecules (proteins or RNA) that sense change through direct interactions with chemical or physical stimuli. In recent years, there have been tremendous advances in understanding the structure and function of these signal receptors, and how interaction with the signal triggers changes in their activity and downstream events. For some systems, this understanding extends to the atomic level. In this unique book, an international team of experts reviews a selection of important model systems, providing a timely snapshot of the current state of research in the field. Sensory Mechanisms in Bacteria opens with an introductory chapter that reviews the diversity of signal recognition mechanisms, illustrating the breadth of the field. Subsequent chapters include descriptions

International Review of Cell and Molecular Biology reviews and details current advances in cell and molecular biology. The IRCMB series has a worldwide readership, maintaining a high standard by publishing invited articles on important and timely topics that are authored by prominent cell and molecular biologists. The articles published in IRCMB have a high impact and an average cited half-life of 9 years. This great resource ranks high amongst scientific journals dealing with cell biology.

In keeping with the broad objectives set for the serial publication of Advances in Structural Biology, Volume 6 contains exhaustive articles from experts in diverse areas of biomedical research. The common thread among the various articles is their relevance to the applications of cell biology to human health.

This volume of "Advances in Cell Aging and Gerontology" critically reviews the rapidly advancing area of telomerase research with a focus at the molecular and cellular levels. The clearly established function of telomerase is to maintain chromosome ends during successive rounds of cell division by adding a six base DNA repeat on to the telomeric ends of chromosomes. As presented in the chapters of this volume, the mechanisms that regulate telomerase expression and activity are complex. Moreover, emerging data suggest additional roles for telomerase in the regulation of cell differentiation and survival.

It is expected that this quite comprehensive volume will provide a valuable resource for graduate students and postdocs in the telomerase field and for established investigators in other fields who are beginning to study telomerase in their particular research program. With an increasing number of proteins being brought into the fold of telomerase research (e.g., DNA damage and repair response proteins, heat-shock proteins, and proteins in various signal transduction cascades) many new scientists are beginning to study this enzyme from novel vantage points.

Marine enzymes and specialized metabolism - Part B, Volume 605 in the Methods in Enzymology series, highlights experimental methods on diverse marine enzymes involved in the construction of bioactive natural product molecules. Unique sections in this new release include discussions on polysaccharide-degrading enzymes from marine gastropods, radical SAM epimerases from sponge microbes, DMS/P demethylase in bacteria, reconstitution of particulate methane monooxygenase into membrane mimetics, the structure and function of cyanobactin enzymes, marine cyanobacterial polyketide beta-branching enzymology, marine cyanobacterial PKS-NRPS enzymology and structural biology, biochemical profiling of DMSP lyases, and more.

The aim of "The Adhesive Interaction of Cells" has been to assemble a series of reviews by leading international experts embracing many of the most important recent developments in this rapidly expanding field. The purpose of all biological research is to understand the form and function of living organisms and, by comprehending the normal, to find explanations and remedies for the abnormal and for disease conditions. The molecules involved in cell adhesion are of fundamental importance to the structure and function of all multicellular organisms. In this book, the contributors focus on the systems of vertebrates, especially mammals, since these are most relevant to human disease. It would have been equally possible to concentrate on developmental processes and adhesion in lower organisms.
A major function of adhesion molecules is to bind cells to each other or to the extracellular matrix, but they are much more than "glue." Adhesions in animal tissues must be dynamic-forming, persisting, or declining in regulated fashion- to facilitate the mobility and turnover of tissue cells. Moreover, the majority of adhesion molecules are transmembrane molecules and thus provide links between the cells and their surroundings. This gives rise to another major function of adhesion molecules, the capacity to transduce signals across the hydrophobic barrier imposed by the plasma membrane. Such signal transduction is crucially important to many aspects of cellular function including the regulation of cell motility, gene expression, and differentiation.
The work in this book progresses through four sections. Part I discusses the four major families of adhesion molecules themselves, the integrins (Green and Humphries), the cadherins (Stappert and Kemler), the selectins (Tedder et al.) and the immunoglobulin superfamily (Simmons); part 2 considers junctional complexes involved in cell interactions: focal adhesions and adherens junctions (Ben Ze'ev), desmosomes (Garrod et al.), and tight junctions (Citi and Cordenonsi). The signaling role of adhesion molecules is the focus of part 3, through integrins and the extracellular matrix (Edwards and Streuli), through platelet adhesion (Du and Ginsberg), and in the nervous system (Hemperley). In part 4, the aim is to show how adhesive phenomena contribute to important aspects of cell behavior and human health. Leukocyte trafficking (Haskard et al.), cancer metastasis (Marshall and Hart), cell migration (Paleck et al.), and implantation and placentation (Damsky et al.) are the topics considered in depth.
The different sections are, of course, not mutually exclusive: it is both undesirable and impossible to separate structure from function when considering cell adhesion. Each chapter has its unique features, but some overlap is both invevitable and valuable since it provides different perspectives on closely related topics. We hope that the whole contributes a valuable and stimulating consideration of this important topic.

Transcriptional Gene Regulation in Health and Disease, Volume 335, the latest release in the International Review of Cell and Molecular Biology reviews and details current advances in cell and molecular biology. The IRCMB series has a worldwide readership, maintaining a high standard by publishing invited articles on important and timely topics that are authored by prominent cell and molecular biologists. The articles published in IRCMB have a high impact and an average cited half-life of nine years. This great resource ranks high amongst scientific journals dealing with cell biology.

Cell Surface GRP78, a New Paradigm in Signal Transduction Biology presents a new paradigm that has emerged in the past decade with the discovery that various intracellular proteins may acquire new functions as cell surface receptors. Two very prominent examples are ATP synthase and GRP78. While the role of cell surface ATP synthase has been reviewed in various books, this book directs its attention to the story of cell surface GRP78.

Mitosis and Meiosis, Part A, Volume 144, a new volume in the Methods in Cell Biology series, continues the legacy of this premier serial with quality chapters authored by leaders in the field. Unique to this updated volume are chapters on Analyzing the Spindle Assembly Checkpoint in human cell culture, an Analysis of CIN, a Functional analysis of the tubulin code in mitosis, Employing CRISPR/Cas9 genome engineering to dissect the molecular requirements for mitosis, Applying the auxin-inducible degradation (AID) system for rapid protein depletion in mammalian cells, Small Molecule Tools in Mitosis Research, Optogenetic control of mitosis with photocaged chemical, and more.

This manual is designed as an intensive introduction to the various tools of molecular biology. It introduces all the basic methods of molecular biology including cloning, PCR, Southern (DNA) blotting, Northern (RNA) blotting, Western blotting, DNA sequencing, oligo-directed mutagenesis, and protein expression.
Key Features
* Provides well-tested experimental protocols for each technique
* Lists the reagents and preparation of each experiment separately
* Contains a complete schedule of experiments and the preparation required
* Includes study questions at the end of each chapter