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."

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.

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 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.

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.

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.

Significant advancements have been made in the study of chromatin structure and function over the past fifty years but none as spectacular as those made in the last decade due to the development of novel techniques and the ability to sequence large stretches of DNA. In Chromatin Protocols, Second Edition, expert researchers delineate these cutting-edge techniques via step-by-step laboratory methods and protocols, which encompass a wide array of topics from the isolation of nucleosomes, assembly of nucleosomes and study of the basic chromatin structure to detailed analysis of histone modifications and chromatin function. Written in the highly successful Methods in Molecular Biology (TM) series style, chapters include brief introductions to the subjects, lists of the necessary materials and reagents, readily reproducible protocols, and Notes sections which highlight tips on troubleshooting and avoiding known pitfalls. Comprehensive and up-to-date, Chromatin Protocols, Second Edition is a valuable tool for scientists studying various aspects of chromatin function and an ideal guide to aid in the development of new techniques as well as new ideas in the field of chromatin biology.

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.

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.

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.

"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.

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.

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.

Bioinformatics is a new and fast expanding area of biology encompassing the organization, analysis and interpretation of the huge amount of data emerging from sequencing and genome projects. Major new programs, software and internet facilities have evolved recently that enable bioinformatic analysis at the whole genome level and more novel technologies are currently being developed. Written by experts in the field, this concise yet informative volume covers all aspects of bioinformatics pertaining to genomic studies. It is an essential book for anyone involved in genomic science or bioinformatics.

This collection of diverse articles by the pioneers of modern genomics takes stock of the current state of the field and elucidates the contribution that sequencing genomes has made to our understanding of microbial metabolism and evolution. Through twenty-eight thought-provoking chapters, the authors describe some of the most common computational methods and their applications to studying pathogenic microorganisms, show how genomics can be used to reconstruct the history and dynamism of the microbial world, and discuss issues as diverse as reconstruction of metabolic pathways, cell cycle processes, microbial evolution, metagenomics, and vaccine development. Additional chapters deal with microarrays and expression analysis and the role of genomic in drug discovery.

Single Cell Diagnostics: Methods and Protocols applies modern
molecular diagnostic techniques to the analysis of single cells, small
numbers of cells, or cell extracts. Emphasis is placed on non-invasive
analysis of single cell metabolites and the direct analysis of RNA and
DNA from single cells, with a focus on polymerase chain reaction and
fluorescence in situ hybridization. The authors, all leading
scientists in the field, provide current and innovative methods for
extracting useful, accurate, and reliable data from single-cell
samples. The stepwise, technically precise protocols presented in this
volume thoroughly describe single-cell assays, from material collection
and isolation of cells to data interpretation. Cell-types analyzed
include embryonic blastomeres, fibroblasts, and lymphocytes. Among the
techniques described are real-time quantitative PCR, isothermal whole
genome amplification, comparative genomic hybridization, real-time
genetic expression analysis, and the production of RNA and cDNA
libraries. This handbook will be essential for researchers and
clinicians, and practitioners providing care for couples seeking
treatment for infertility or preimplantation genetic diagnosis.

Structural Genomics and Drug Discovery: Methods and Protocols focuses on high throughput structure determination methods and how they can be applied to lay the groundwork for structure aided drug discovery. The methods and protocols that are described can be applied in any laboratory interested in using detailed structural information to advance the initial stages of drug discovery. 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, Structural Genomics and Drug Discovery: Methods and Protocols seeks to aid scientists in the further study into structural genomics approach as an efficient initial step toward drug discovery and the methods described will be useful to anyone interested in moving in this direction.

Several developmental and historical threads are woven and displayed in these two volumes of Bacterial Artificial Chromosomes, the first on Library Construction, Physical Mapping, and Sequencing, and the second on Fu- tional Studies. The use of large-insert clone libraries is the unifying feature, with many diverse contributions. The editors have had quite distinct roles. Shaying Zhao has managed several BAC end-sequencing projects. Marvin Stodolsky during 1970-1980 contributed to the elucidation of the natural b- teriophage/prophage P1 vector system. Later, he became a member of the Genome Task Group of the Department of Energy (DOE), through which s- port flowed for most clone library resources of the Human Genome Program (HGP). Some important historical contributions are not represented in this volume. This preface in part serves to mention these contributions and also briefly surveys historical developments. Leon Rosner (deceased) contributed substantially in developing a PAC library for drosophila that utilized a PI virion-based encapsidation and tra- fection process. This library served prominently in the Drosophila Genome Project collaboration. PACs proved easy to purify so that they substantially replaced the YACs used earlier. Much of the early automation for massive clone picking and processing was developed at the collaborating Lawrence Berkeley National Laboratory. However, the P1 virion encapsidation system itself was too fastidious, and P1 virion-based methods did not gain popularity in other genome projects.

Biological rhythms, such as the sleep-wake cycle or circadian clock, are an intriguing aspect of biology. The regulation of daily rhythmicity has long been a mystery, up until the mid-1980's when a key gene in the fruitfly, "Drosophila melanogaster," was molecularly identified. Genetic and molecular chronobiology of "Drosophila" has been a driving force in this field of inquiry ever since. This book describes and evaluates all of the studies of this sort, discussing the manner by which these investigations have spread out in various directions of rhythmic biology, including genetic and molecular approaches used on other insect species.
* Discusses rhythm genetics in insects, from early investigations to current state-of-the-art
* Presents all relevant mutants and genes
* Highlights the mystery of the "clock mechanism" in full detail including the remaining puzzles to be solved

"Your genome is an email attachment" What a difference a few years can make? In 2001, to a global fanfare, the completion of the frst draft sequence of the human genome was announced. This had been a Herculean effort, involving thousands of researchers and millions of dollars. Today, a project to re-sequence 1,000 genomes is well underway, and within a year or two, your own "personal genome" is likely to be available for a few thousand pounds, a price that will undoubtedly decrease further. We are fast approaching the day when your genome will be available as an email attachment (about 4 Mb). The key to this feat is the fact that any two human genomes are more than 99% identical, so rather than representing every base, there is really only a requirement to store the 1% of variable sequence judged against a common reference genome. This brings us directly to the focus of this edition of Methods in Molecular Biology, Genetic Variation. The human genome was once the focus of biology, but now individual genome var- tion is taking the center stage. This new focus on individual variation ultimately democ- tizes biology, offering individuals insight into their own phenotype. But these advances also raise huge concerns of data misuse, misinterpretation, and misunderstanding. The immediacy of individual genomes also serves to highlight our relative ignorance of human genetic variation, underlining the need for more studies of the nature and impact of genetic variation on human phenotypes.

MicroRNA Protocols provides diverse, novel, and useful descriptions of miRNAs in several species, including plants, worms, flies, fish, chicks, mice, and humans. These include some useful adaptations and applications that could be relevant to the wider research community who are already familiar with the identification of miRNAs. This volume will stimulate the reader to explore diverse ways to understanding the mechanism in which miRNAs facilitate the molecular aspects of the biomedical research.

Eukaryotic DNA Damage Surveillance and Repair contains chapters from experts in the field of DNA damage detection, repair, and cell cycle control. The work reviews current understanding of how different types of DNA damage are detected and focuses on how these surveillance mechanisms are coupled to processes of DNA repair, cell cycle control, and apoptosis.
The title will be of interest to undergraduate/postgraduate students and academics alike.

RNA interference (RNAi), in which RNA silences RNA, is the most recent discovery to revolutionize the study of biology. In RNAi: Design and Applications, leaders in the field contribute state-of-the-art, easy to follow methods and bench protocols designed for practical, everyday use of RNAi in biological research. Divided into two parts, this comprehensive volume covers fundamentals including designs of RNAi, biochemical assay protocols for the major components of RNAi, and study of potential off-target effects, followed by an extensive section covering various applications of RNAi in diverse model organisms and systems, from antiviral and anticancer applications to altering flower color in plants. Cutting edge and clearly written, RNAi: Design and Applications enables a researcher with standard molecular biological training to perform major RNAi-related experiments and contribute to this revolutionary, growing field.