DNA Amplification


Book Description

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.




PCR Technology


Book Description

This is an introduction to the methods and applications of polymerase chain reaction (PCR) technology, a technology developed by Erlich's group at Cetus and Cetus, and is expected to be used in all biology laboratories worldwide within the next few years.




Methods in DNA Amplification


Book Description

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.




Modern Applications of DNA Amplification Techniques


Book Description

In the ten years since the first publication on PCR (Saiki et al. , 1985), this in vitro method of nucleic acid replication and modification has grown to rival in popularity traditional microbiological, genetical und technical procedures for cloning, sequencing, gene detecting and related procedures. To date the PCR literature has emphasized six main areas of application: genetic mapping, detection of mutations, genetic polymorphism, transcriptional splicing and regulation, molecular virology and quantitative procedures. The overwhelming focus of quantification of DNA or RNA by PCR has been on human microbiology and oncological problems. The exquisite sensitivity of PCR gives this method the ability to detect extremely rare DNAs, mRNAs, mRNAs in small numbers of cells or in small amounts of tissue, and mRNAs expressed in mixed-cell populations. However, the exact and accurate quantification of specific nucleic acids in biological samples is in spite of numerous publications in that field still a general problem: during the peR process, an unknown initial number of target sequences are used as a template from which a large quantity of specific product can be obtained. Although the amount of product formed is easy to determine, it is difficult to deduce the initial copy number of the target molecule because the efficiency of the peR is largely unknown.




The Polymerase Chain Reaction


Book Description

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, " ...




Molecular Techniques in Taxonomy


Book Description

Taxonomy is fundamental to understanding the variety of life forms, and exciting expansions in molecular biology are re- volutionising the obtained data. This volume reviews the ma- jor molecular biological techniques that are applied in ta- xonomy. The chapters are arranged in three main sections:1) Overviews of important topics in molecular taxonomy; 2) Case studies of the successful application of molecular methods to taxonomic and evolutionary questions; 3) Protocols for a range of generally applicable methods. The described techni- ques include DNA-DNA hybridization, DNA fingerprinting, RFLP analysis, and PCR sequencing.




PCR Technology


Book Description

Polymerase chain reaction (PCR) technology is a revolutionary innovation which enables scientists to rapidly generate large amounts of genetic material from a slight trace which would otherwise be too small to analyze. With applications in both research and diagnostics, PCR is becoming a standard procedure in biotechnology and medical diagnostic laboratories. This book is an introduction and guide to the new technology, covering the basic methodologies and their applications in research and medicine, emphasizing practical aspects. Each chapter is written by pioneers in the field and most include detailed protocols and favorite PCR "recipes". Students and researchers in all areas of biotechnology and molecular biology will find this book the introduction to PCR they've been looking for.




Principles and Technical Aspects of PCR Amplification


Book Description

Kary Mullis was awarded a Nobel Prize for inventing the PCR technique more than a decade ago in 1993. Since its "discovery", multiple adaptations and variations of the standard PCR technique have been described. This publication aims to provide the reader with a guide to the standard PCR technique and its many available variants, with particular emphasis being placed on the role of these PCR techniques in the clinical diagnostic laboratory (the central theme of this book).




Nucleic Acid Amplification Technologies


Book Description

Providing current information and guidance on the uses of various nucleic acid amplification technologies for clinical laboratory diagnosis, this book goes beyond the Polymerase Chain Reaction to explore a broader range of important alternative DNA/RNA amplification methods including the Ligase Chain Reaction, Q[beta] Replicase Assays and TMA. There are many examples of specific applications of these technologies, discussions of yet unresolved issues and demonstrations of the relevance of these technologies to medical research and disease diagnostics. Individual chapters cover uses of these methods in clinical situations such as detection of food pathogens, viral infections, STDs, Mycobacteria drug resistance mutations, and heritable diseases. Automation, diagnostic test evaluation, and the synthesis of artificial DNA are also discussed. This book is designed for all biomedical scientists interested in the application of molecular biology to clinical diagnosis.




Encyclopedia of Astrobiology


Book Description

The interdisciplinary field of Astrobiology constitutes a joint arena where provocative discoveries are coalescing concerning, e.g. the prevalence of exoplanets, the diversity and hardiness of life, and its increasingly likely chances for its emergence. Biologists, astrophysicists, biochemists, geoscientists and space scientists share this exciting mission of revealing the origin and commonality of life in the Universe. The members of the different disciplines are used to their own terminology and technical language. In the interdisciplinary environment many terms either have redundant meanings or are completely unfamiliar to members of other disciplines. The Encyclopedia of Astrobiology serves as the key to a common understanding. Each new or experienced researcher and graduate student in adjacent fields of astrobiology will appreciate this reference work in the quest to understand the big picture. The carefully selected group of active researchers contributing to this work and the expert field editors intend for their contributions, from an internationally comprehensive perspective, to accelerate the interdisciplinary advance of astrobiology.