Beginning with the Escherichia coli co protein, or bacterial DNA topoisomerase I, an ever-increasing number of enzymes has been identified that catalyze changes in the linkage of DNA strands. DNA topoisomerases are ubiquitous in nature and have been shown to play critical roles in most p- cesses involving DNA, including DNA replication, transcription, and rec- bination. These enzymes further constitute the cellular targets of a number of clinically important antibacterial and anticancer agents. Thus, further studies of DNA topology and DNA topoisomerases are critical to advance our und- standing of the basic biological processes required for cell cycle progression, cell division, genomic stability, and development. In addition, these studies will continue to provide critical insights into the cytofoxic action of drugs that target DNA topoisomerases. Such mechanistic studies have already played an important role in the development and clinical application of antimicrobial and chemotherapeutic agents. The two volumes of DNA Topoisomerase Protocols are designed to help new and established researchers investigate all aspects of DNA topology and the function of these enzymes. The chapters are written by prominent investigators in the field and provide detailed background information and st- by-step experimental protocols. The topics covered in Volume I; DNA Topology and Enzymes, range from detailed methods to analyze various aspects of DNA structure, from linking number, knotting/unknotting, site-specific recombi- tion, and decatenation to the overexpression and purification of bacterial and eukaryotic DNA topoisomerases from a variety of cell systems and tissues.

A comprehensive treatment of the characterisation techniques used in investigating inorganic and organic molecules that interact with biomolecules is presented to the reader in a clear fashion. The work consists of two parts: (i) synthetic aspects of metallointercalators along with targeting and improving transport and (ii) the various techniques that are used for probing their interactions, such as; DNA-NMR, PGSE-NMR, DNA ESI-MS, Linear and Circular Dichroism, Fluorescence Spectroscopy, Confocal Microscopy, Viscosity, TGA and dialysis, Microarrays, biological analysis. Chapters are devoted to the synthesis and the techniques used to study the interactions of inorganic complexes with biomolecules. Considerably detailed examples are used to help illustrate the application of these techniques. This book is a useful resource for an array of inorganic and organic advanced undergraduate and graduate courses and for researchers in drug discovery.

Damage to DNA by both exogenous and endogenous sources is increasingly regarded as highly important in the initiation and progression of cancer and in the occurance of other pathological events. DNA damage caused by reactive oxygen-derived species, also called oxidative DNA damage, is most the frequent type encountered by aerobic cells. Mechanistic studies of carcinogenesis indicate an important role of this type of damage to DNA. There is also strong evidence to support the role of oxidative DNA damage in the aging process. DNA damage is opposed in vivo by repair systems. If not repaired, DNA damage may lead to detrimental biological consequences. Therefore, the repair of DNA damage is regarded as one of the essential events in all life forms. In recent years the field of DNA repair has flourished due to new findings on DNA repair mechanisms and the molecular basis of cancer. A detailed knowledge of mechanisms of DNA damage and repair, and how individual repair enzymes function may lead to manipulation of DNA repair in cells and ultimately to an increase of the resistence of human cells to DNA-damaging agents. This volume covers the most recent devlopments in this research field and contains contributions from scientists working in the fields of biochemistry, molecular biology, enzymology, biomedical science, and radiation biology.

Ancient DNA Typing is a comprehensive manual about the analysis of ancient and degraded DNA. It is an equally useful textbook and working tool for various disciplines such as: Anthropology, Archaeology, Forensic Medicine, Conservation Biology, Environmental Sciences and Food Engineering.The reader benefits from a clear division of strategies, methods and applications with many interesting examples of the analysis of historic skeletal and cultural remains. The book contains a detailed protocol chapter which serves as a lab manual with the most important methods for DNA extraction, PCR and electrophoretic analysis of degraded samples. The appendix helps to set up a PCR lab specializing in ancient DNA research and answers the most frequent questions concerning technical problems in investigations of degraded DNA.

Genetic recombination, in the broadest sense, can be defined as any process in which DNA sequences interact and undergo a transfer of information, producing new "recombinant" sequences that contain information from each of the original molecules. All organisms have the ability to carry out recombination, and this striking universality speaks to the essential role recombination plays in a variety of biological processes fundamentally important to the maintenance of life. Such processes include DNA repair, regulation of gene expression, disease etiology, meiotic chromosome segregation, and evolution. One important aspect of recombination is that it typically occurs only between sequences that display a high degree of sequence identity. The stringent requirement for homology helps to ensure that, under normal circumstances, a cell is protected from deleterious rearrangements since a swap of genetic information between two nearly identical sequences is not expected to dramatically alter a genome. Recombination between dissimilar sequences, which does happen on occasion, may have such harmful consequences as chromosomal translocations, deletions, or inversions. For many organisms, it is also important that recombination rates are not too high lest the genome become destabilized. Curiously, certain organisms, such as the trypanosome parasite, actually use a high rate of recombination at a particular locus in order to switch antigen expression continually and evade the host immune system effectively.

Humanity's physical design flaws have long been apparent--we get hemorrhoids and impacted wisdom teeth, for instance--but do the imperfections extend down to the level of our genes? Inside the Human Genome is the first book to examine the philosophical question of why, from the perspectives of biochemistry and molecular genetics, flaws exist in the biological world. Distinguished evolutionary geneticist John Avise offers a panoramic yet penetrating exploration of the many gross deficiencies in human DNA--ranging from mutational defects to built-in design faults--while at the same time offering a comprehensive treatment of recent findings about the human genome. The author shows that the overwhelming scientific evidence for genomic imperfection provides a compelling counterargument to intelligent design. He also develops a case that theologians should welcome rather than disavow these discoveries. The evolutionary sciences can help mainstream religions escape the shackles of Intelligent Design, and thereby return religion to its rightful realm--not as the secular interpreter of the biological minutiae of our physical existence, but rather as a respectable philosophical counselor on grander matters of ultimate concern.

Protocols for Nucleic Acid Analysis by Non-radioactive Probes, Second Edition provides a firm background on the basic preparative protocols required for the analysis of nucleic acids by nonradioactive methods. Presenting the methodologies using amazing new applications, this volume offers guide chapters on nucleic acid extractions, preparation of nucleic acid blots, and labeling of nucleic acids with nonradioactive haptens. New fluorescent techniques such as Real Time PCR and microarrays are also included, allowing users to get a nonradioactive protocol implemented in the laboratory with minimum adaptation required and fastest time to results.
The protocols follow the successful Methods in Molecular Biology series format, each offering step-by-step laboratory instructions, an introduction outlining the principles behind the technique, lists of the necessary equipment and reagents, and tips on troubleshooting and avoiding known pitfalls.

Over the last decade Life Science has undergone an accelerated evolution, culminating in the -omics era characterized by the development of a multitude of high throughput methods that are becoming more routinely applied in biochemistry labs. In Functional Genomics: Methods and Protocols, Second Edition expert researchers in the field detail many of the methods which are now commonly used for studies in the life sciences focusing on the dynamic aspects of the transcriptome, proteome and metabolome, respectively.Written in the highly successful Methods in Molecular Biology (TM) 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, Functional Genomics: Methods and Protocols, Second Edition seeks to aid scientists in establishing or extending technologies and techniques in their laboratories.

This book examines the increasing significance of DNA profiling for crime investigation in modern society. It focuses on developments in the UK as the world-leader in the development and application of forensic DNA technology, and in the construction of DNA databases as an essential element in the successful use of DNA for forensic purposes. The book uses data collected from funded research into police uses of the UK National DNA Database (NDNAD) to describe the relationship between scientific knowledge and police investigations. It refers to some of the major UK criminal cases in which DNA evidence has been presented and contested. Chapters in the book explain the scientific developments which have enabled DNA profiling to be applied to criminal investigation, the ways in which the state has directed this, and how genetic technology has risen to such preeminence; how DNA evidence moved from its use in individual prosecutions to a major role in intelligence led policing, and saw the de

In the past few years, antisense methodology has moved from in vitro studies to in vivo studies and first human trials. While the basic concept of antisense technology is simple, the methodological problems associated with its use are numerous and complex. Antisense- based methods have proven to be a field of research where careful attention to experimental protocols and appropriate controls is necessary. The Manual of Antisense Methodology emphasizes the application of antisense oligonucleotides, and is a guide for the identification of antisense and non-antisense effects in different experimental settings. The work is organized into three sections: antisense application in vitro, antisense application in vivo (animal models) and finally, clinical antisense studies. Where at all possible, the methods are described in sufficient detail to allow reproduction of a given experiment. The Manual of Antisense Methodology will be of interest to researchers in immunology, cancer research, pharmacology and internal medicine; and physicians conducting clinical studies in these fields.

Y.A. Berlin, I.V. Kurnikow, D. Beratan, M.A. Ratner, A.L. Burin: DNA Electron Transfer Processes: Some Theoretical Notions; N. Rosch, A.A. Voityuk: Quantum Chemical Calculation of Donor-Acceptor Coupling for Charge Transfer in DNA; E. Conwell: Polarons and Transport in DNA; Z. Cai, M.D.

This volume is divided in four sections; covering genome wide approaches, techniques for characterize of paRNA structural features are described, selecting pa-RNA, and paRNA therapeutic potential. Chapters describe how siRNAsdirected against paRNAs can be applied in vivo to modulate transcription of important genes controlled by paRNAs. 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. Authoritative and cutting-edge, Promoter Associated RNA: Methods and Protocols aims to demonstrate paRNAs as new class of regulatory molecules, to further investigate and value as tools for fine transcriptional tuning.

Quantitative trait locus (QTL) mapping is used to discover the genetic and molecular architecture underlying complex quantitative traits. It has important applications in agricultural, evolutionary, and biomedical research. R/qtl is an extensible, interactive environment for QTL mapping in experimental crosses. It is implemented as a package for the widely used open source statistical software R and contains a diverse array of QTL mapping methods, diagnostic tools for ensuring high-quality data, and facilities for the fit and exploration of multiple-QTL models, including QTL x QTL and QTL x environment interactions. This book is a comprehensive guide to the practice of QTL mapping and the use of R/qtl, including study design, data import and simulation, data diagnostics, interval mapping and generalizations, two-dimensional genome scans, and the consideration of complex multiple-QTL models. Two moderately challenging case studies illustrate QTL analysis in its entirety.

The book alternates between QTL mapping theory and examples illustrating the use of R/qtl. Novice readers will find detailed explanations of the important statistical concepts and, through the extensive software illustrations, will be able to apply these concepts in their own research. Experienced readers will find details on the underlying algorithms and the implementation of extensions to R/qtl. There are 150 figures, including 90 in full color.

Introduction The goal of this book is to introduce XML to a bioinformatics audience. It does so by introducing the fundamentals of XML, Document Type De?nitions (DTDs), XML Namespaces, XML Schema, and XML parsing, and illustrating these concepts with speci?c bioinformatics case studies. The book does not assume any previous knowledge of XML and is geared toward those who want a solid introduction to fundamental XML concepts. The book is divided into nine chapters: Chapter 1: Introduction to XML for Bioinformatics. This chapter provides an introduction to XML and describes the use of XML in biological data exchange. A bird's-eye view of our ?rst case study, the Distributed Annotation System (DAS), is provided and we examine a sample DAS XML document. The chapter concludes with a discussion of the pros and cons of using XML in bioinformatic applications. Chapter 2: Fundamentals of XML and BSML. This chapter introduces the fundamental concepts of XML and the Bioinformatic Sequence Markup Language (BSML). We explore the origins of XML, de?ne basic rules for XML document structure, and introduce XML Na- spaces. We also explore several sample BSML documents and visualize these documents in the TM Rescentris Genomic Workspace Viewer.

The recent progress achieved in aquatic genomics research has important implications in science and aquaculture. Researchers' success in genome sequencing of the fugu has provided a foundation not only for comparative fish genomics but also for genetic studies of other vertebrates. This volume, with contributions by leading scientists from around the world, provides the latest information on genomic studies of a variety of aquatic organisms including fish, crustaceans, and shellfish, with descriptions of such methodologies as genome analysis, gene mapping, DNA markers, and EST analysis. Other topics include regulation of gene expression, stress and immune responses, sex differentiation, hormonal control, and transgenic fish. This book is a valuable resource for all scientists whose work involves aquatic organisms, especially for those in the fields of aquatic genomics, fish immunology, and aquatic biotechnology.

The primary purpose of this volume is to demonstrate the range of applications of the Pyrosequencing technology in research and diagnostics and to provide detailed protocols. Beginning with an up-to-date overview of the biochemistry, the volume continues with quantitative analysis of genetic variation, ratio of expressed alleles at the RNA level, analysis of DNA methylation, global DNA methylation assays, specialized applications for DNA methylation analysis including loss of imprinting, single blastocyst analysis, allele-specific DNA methylation patterns, DNA methylation patterns associated with specific histone modifications. The volume further details tools and protocols for the detection of viruses and bacteria, and genetic and epigenetic analyses for forensics using Pyrosequencing. As a volume in the highly successful Methods in Molecular Biology series, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols and tips on troubleshooting and avoiding known pitfalls. Comprehensive and adaptable, Pyrosequencing: Methods and Protocols, Second Edition will greatly aid doctorial students, postdoctoral investigators and research scientists studying different aspects of genetics and cellular and molecular biology.

Genome Exploitation: Data Mining the Genome is developed from the 23rd Stadler Genetic Symposium. This volume discusses and illustrates how scientists are going to characterize and make use of the massive amount of information being accumulated about the plant and animal genomes. Genome Exploitation: Data Mining the Genome is a state-of-the-art picture on mining the Genome databases. This is one of the few times that researchers in both plants and animals will be working together to create a seminal data resource.

This comprehensive yet balanced work emphasizes the principles and rationale underlying recombinant DNA methodology while furnishing a general understanding of the experimental protocols-suggesting flexible approaches to resolving particular molecular necessities that are easily adaptable to readers' specific applications. Features summary tables presenting at-a-glance information on practices of recombinant DNA methodologies! Recombinant DNA Principles and Methodologies discusses basic and advanced topics requisite to the employment of recombinant DNA technology, such as -plasmid biology -nucleic acid biochemistry -restriction enzymes -cloning strategies -gel electrophoresis -southern and northern blotting -preparation of probes -phage lambda biology -cosmids and genome analysis -cloned gene expression -polymerase chain reaction -conventional and automated DNA sequencing -site-directed mutagenesis -and more! Elucidating the material with over 2250 edifying references, equations, drawings, and photographs, this state-of-the-art resource is a valuable hands-on guide for molecular and cell biologists, biochemists, bioprocess technologists, applied and industrial microbiologists, virologists, geneticists, chemical engineers, and upper-level undergraduate and graduate students in these disciplines.

In this volume, internationally recognized 'Neurospora' experts critically review the most important research and demonstrate the breadth of applications to industrial biology, biofuels, agriculture, and human health.

Environmental DNA (eDNA) refers to DNA that can be extracted from environmental samples (such as soil, water, feces, or air) without the prior isolation of any target organism. The analysis of environmental DNA has the potential of providing high-throughput information on taxa and functional genes in a given environment, and is easily amenable to the study of both aquatic and terrestrial ecosystems. It can provide an understanding of past or present biological communities as well as their trophic relationships, and can thus offer useful insights into ecosystem functioning. There is now a rapidly-growing interest amongst biologists in applying analysis of environmental DNA to their own research. However, good practices and protocols dealing with environmental DNA are currently widely dispersed across numerous papers, with many of them presenting only preliminary results and using a diversity of methods. In this context, the principal objective of this practical handbook is to provide biologists (both students and researchers) with the scientific background necessary to assist with the understanding and implementation of best practices and analyses based on environmental DNA.

Recent progress in high-throughput technologies and genome wide transcriptome studies have lead to a significant scientific milestone of discovering non-coding RNAs (ncRNAs) which spans through a major portion of the genome. These RNAs most often act as riboregulators, and actively participate in the regulation of important cellular functions at the transcriptional and/or post-transcriptional levels rather than simply being an intermediated messenger between DNA and proteins. As the appreciation for the importance of ncRNAs continues to emerge, it is also increasingly clear that these play critical roles in gene regulatory processes during development and differentiation. Further, regulatory RNAs are useful biomarkers for diagnosis of diseases. Hence these RNA regulators are essential to the development of therapeutics. This book on "Regulatory RNAs" offers a comprehensive view on our current understanding of these regulatory RNAs viz. siRNA, miRNA, piRNA, snoRNA, long non-coding RNA, small RNA etc. It addresses both the biogenesis and mechanism of action of regulatory RNAs with a primary focus on their annotation, experimental methodologies (microarray, next-gen sequencing etc.) for their discovery, computational tools for their prediction, and above all, applications of these revolutionary regulatory molecules in understanding biological systems and diseases, including therapeutics. This comprehensive volume is intended for readers with research or teaching interests in ncRNA biology and will provide a major information resource on current research in the fast-moving fields of RNA and gene expression regulation. Cutting-edge and concise, "Regulatory RNAs: Basics, Methods and Applications" promises to support vital research in the field of regulatory RNAs, ever-continuing to grow rapidly and gain increasing importance in basic and translational biology.

Understanding of the molecular basis of DNA damage and its repair has increased dramatically in recent years and there is now a substantial knowledge concerning the products that arise from free-radical attack on DNA. Free-radical DNA damage may lead to mutations, cancer and cell death. There are various free radical sources, notably ionizing radiation or oxidative stress. In the radiotherapy of cancer and in the case of some anticancer drugs, use is made of cell killing by excessive DNA damage.nbsp; The mechanisms that lead to these products have been studied on the model level and with small double-stranded DNA fragments and are discussed in some detail. The basics of the underlying free-radical chemistry are dealt with in separate chapters. Understanding of the molecular basis of DNA damage and its repair has increased dramatically in recent years, and substantial knowledge now exists concerning the products arising from free-radical attack on DNA. Free-radical DNA damage may lead to mutations, cancer, and cell death. Free radicals have various sources, notably ionizing radiation and oxidative stress. In radiotherapy for cancer and with some anticancer drugs, use is made of cell death by excessive DNA damage. The mechanisms leading to products of free-radical attack which have been studied in models and with small double-stranded DNA fragments are discussed in detail, and the basics of the underlying free-radical chemistry are dealt with in separate chapters.

Cases in Laboratory Genetics and Genomics (LGG) Practice instructs readers in lab-based diagnosis of genetic conditions, including inborn and acquired disorders, using cytogenetics and molecular genetics technologies. This entirely case-based book covers a wide range of genetic cases from prenatal to postnatal and oncology genetic disorders, which lab professionals and geneticists encounter daily in the diagnostic field. Each disorder discussed includes a section on clinical background, clinical indication, tests ordered, laboratory tests performed, test results, results with interpretations, future testing and recommendations, and references. The book will help lab professionals understand and navigate clinical cases using an integrative approach, and thoroughly understand the methodologies and interpretations involved in high complexity genetic testing.