Whereas most books on DNA amplification focus on PCR-based technologies, this volume presents a wider range of methods to amplify DNA with an emphasis on their diverse applications. The book covers both well-established and newly-developed protocols including ligation-based thermocycling approaches, real-time PCR and other new PCR developments, plus several powerful non-PCR isothermal DNA amplification techniques, for example: real-time strand displacement amplification (SDA), rolling-circle amplification (RCA) and multiple-displacement amplification (MDA). An entire section is devoted to a group of enzymes, both natural and engineered, which are employed for DNA amplification and related purposes. In addition, the use of DNA amplification in the detection of non-DNA analytes is presented.
This book offers an overview of state-of-the-art in non amplified DNA detection methods and provides chemists, biochemists, biotechnologists and material scientists with an introduction to these methods. In fact all these fields have dedicated resources to the problem of nucleic acid detection, each contributing with their own specific methods and concepts. This book will explain the basic principles of the different non amplified DNA detection methods available, highlighting their respective advantages and limitations. Non-amplified DNA detection can be achieved by adopting different techniques. Such techniques have allowed the commercialization of innovative platforms for DNA detection that are expected to break into the DNA diagnostics market. The enhanced sensitivity required for the detection of non amplified genomic DNA has prompted new strategies that can achieve ultrasensitivity by combining specific materials with specific detection tools. Advanced materials play multiple roles in ultrasensitive detection. Optical and electrochemical detection tools are among the most widely investigated to analyze non amplified nucleic acids. Biosensors based on piezoelectric crystal have been also used to detect unamplified genomic DNA. The main scientific topics related to DNA diagnostics are discussed by an outstanding set of authors with proven experience in this field.
The polymerase chain reaction (PCR) - an in Vitro techniques for producing large amounts of a specific DNA fragment - has rapidly become established as one of the most important, impressive and fascinating methods of molecular biology as well as clinical diagnostics. In the seven years since'the technique was published, it has had a major impact on medical research. However, as there are still problems in instruments, standardized protocols for diagnostic applications and unsolved difficulties to avoid cross-contaminations on the one hand and on the other hand the even present question of how to interpret the biological value of a PCR result, most clinicians prefer to further wait until these topics are clarified. It is the aim of this book to give the reader lab-proven protocols from experienced scientists as well as a general introduction to alternative DNA-amplification procedures and their possible usage such as the NASBA or LCR. This book is divided into four major parts to provide a theoretical (first and second section) and a practical framework for a better understanding of the new technology. In the first part we provide an up-to-date summary of basic problems in this rapidly evolving field. We demonstrate, for example how to use fixed tissue materials and how to quantify PCR products as well as how to prepare nucleic acids in a safe, convenient and proper way, or even how to sequence directly PCR products for the analysis of the DNA structure.
Holland-Frei Cancer Medicine, Ninth Edition, offers a balanced view of the most current knowledge of cancer science and clinical oncology practice. This all-new edition is the consummate reference source for medical oncologists, radiation oncologists, internists, surgical oncologists, and others who treat cancer patients. A translational perspective throughout, integrating cancer biology with cancer management providing an in depth understanding of the disease An emphasis on multidisciplinary, research-driven patient care to improve outcomes and optimal use of all appropriate therapies Cutting-edge coverage of personalized cancer care, including molecular diagnostics and therapeutics Concise, readable, clinically relevant text with algorithms, guidelines and insight into the use of both conventional and novel drugs Includes free access to the Wiley Digital Edition providing search across the book, the full reference list with web links, illustrations and photographs, and post-publication updates
Whole genome amplification generates microgram quantities of genomic DNA starting from a sample of as little as a few femtograms and so is a vital technique when sample material is limited, as well as for high-throughput assays. Whole Genome Amplification: Methods Expressis a comprehensive up-to-date laboratory manual for this key technique. It provides detailed step-by-step protocols as well as hints and tips for success and troubleshooting, taking readers through all aspects of whole genome amplification. This book is an essential practical guide for any researcher currently using PCR for genomic amplification or who wishes to do so in future.
1Bimal D. Theophilus and Ralph Rapley provide biological and clinical investigators with a comprehensive collection of new, recent, and updated PCR-based screening methods suitable for detecting the presence of both known and novel mutations. The methods cover point mutations (e.g., ASO-PCR, SSCP, DGGE, chemical cleavage), deletions (multiplex PCR, FISH, blotting), non-sense mutations (PTT), and more. The new and exciting techniques of DNA array analysis, along with such recently developed experimental methods as conformation-sensitive gel electrophoresis, are also included. Each chapter explains the basic theory behind the technique and provides valuable notes essential for its successful execution.
Metallic nanoparticles display fascinating properties that are quite different from those of individual atoms, surfaces or bulk rmaterials. They are a focus of interest for fundamental science and, because of their huge potential in nanotechnology, they are the subject of intense research effort in a range of disciplines. Applications, or potential applications, are diverse and interdisciplinary. They include, for example, use in biochemistry, in catalysis and as chemical and biological sensors, as systems for nanoelectronics and nanostructured magnetism (e.g. data storage devices), where the drive for further miniaturization provides tremendous technological challenges and, in medicine, there is interest in their potential as agents for drug delivery.The book describes the structure of metallic nanoparticles, the experimental and theoretical techniques by which this is determined, and the models employed to facilitate understanding. The various methods for the production of nanoparticles are outlined. It surveys the properties of clusters and the methods of characterisation, such as photoionization, optical spectroscopy, chemical reactivity and magnetic behaviour, and discusses element-specific information that can be extracted by synchrotron-based techniques such as EXAFS, XMCD and XMLD. The properties of clusters can vary depending on whether they are free, deposited on a surface or embedded in a matrix of another material; these issues are explored. Clusters on a surface can be formed by the diffusion and aggregation of atoms; ways of modelling these processes are described. Finally we look at nanotechnology and examine the science behind the potential of metallic nanoparticles in chemical synthesis, catalysis, the magnetic separation of biomolecules, the detection of DNA, the controlled release of molecules and their relevance to data storage.The book addresses a wide audience. There was a huge development of the subject beginning in the mid-1980s where researchers began to study the properties of free nanoparticle and models were developed to describe the observations. The newcomer is introduced to the established models and techniques of the field without the need to refer to other sources to make the material accessible. It then takes the reader through to the latest research and provides a comprehensive list of references for those who wish to pursue particular aspects in more detail. It will also be an invaluable handbook for the expert in a particular aspect of nanoscale research who wishes to acquire knowledge of other areas.The authors are specialists in different aspects of the subject with expertise in physics and chemistry, experimental techniques and computational modelling, and in interdisciplinary research. They have collaborated in research. They have also collaborated in writing this book, with the aim from the outset of making it is a coherent whole rather than a series of independent loosely connected articles.* Appeals to a wide audience* Provides an introduction to established models and techniques in the field* Comprehensive list of references
Accompanying CD-ROM contains ... "a companion eBook version of Molecular diagnostics : for the clinical laboratorian, Second edition ... for downloading and use in the reader's PC or PDA."--Page 4 of cover.
James D. Watson When, in late March of 1953, Francis Crick and I came to write the first Nature paper describing the double helical structure of the DNA molecule, Francis had wanted to include a lengthy discussion of the genetic implications of a molecule whose struc ture we had divined from a minimum of experimental data and on theoretical argu ments based on physical principles. But I felt that this might be tempting fate, given that we had not yet seen the detailed evidence from King's College. Nevertheless, we reached a compromise and decided to include a sentence that pointed to the biological significance of the molecule's key feature-the complementary pairing of the bases. "It has not escaped our notice," Francis wrote, "that the specific pairing that we have postulated immediately suggests a possible copying mechanism for the genetic material." By May, when we were writing the second Nature paper, I was more confident that the proposed structure was at the very least substantially correct, so that this second paper contains a discussion of molecular self-duplication using templates or molds. We pointed out that, as a consequence of base pairing, a DNA molecule has two chains that are complementary to each other. Each chain could then act ". . . as a template for the formation on itself of a new companion chain, so that eventually we shall have two pairs of chains, where we only had one before" and, moreover, " ...
