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.




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, " ...




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.




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.







DNA Technology in Forensic Science


Book Description

Matching DNA samples from crime scenes and suspects is rapidly becoming a key source of evidence for use in our justice system. DNA Technology in Forensic Science offers recommendations for resolving crucial questions that are emerging as DNA typing becomes more widespread. The volume addresses key issues: Quality and reliability in DNA typing, including the introduction of new technologies, problems of standardization, and approaches to certification. DNA typing in the courtroom, including issues of population genetics, levels of understanding among judges and juries, and admissibility. Societal issues, such as privacy of DNA data, storage of samples and data, and the rights of defendants to quality testing technology. Combining this original volume with the new update-The Evaluation of Forensic DNA Evidence-provides the complete, up-to-date picture of this highly important and visible topic. This volume offers important guidance to anyone working with this emerging law enforcement tool: policymakers, specialists in criminal law, forensic scientists, geneticists, researchers, faculty, and students.




PCR Protocols


Book Description

The correct procedures you need for frustration-free PCR methods and applications are contained in this complete, step-by-step, clearly written, inexpensive manual. - Avoid contamination--with specific instructions on setting up your lab - Avoid cumbersome molecular biological techniques - Discover new applications




Advanced Techniques in Diagnostic Microbiology


Book Description

Clinical microbiologists are engaged in the field of diagnostic microbiology to determine whether pathogenic microorganisms are present in clinical specimens collected from patients with suspected infections. If microorganisms are found, these are identified and susceptibility profiles, when indicated, are determined. During the past two decades, technical advances in the field of diagnostic microbiology have made constant and enormous progress in various areas, including bacteriology, mycology, mycobacteriology, parasitology, and virology. The diagnostic capabilities of modern clinical microbiology laboratories have improved rapidly and have expanded greatly due to a technological revolution in molecular aspects of microbiology and immunology. In particular, rapid techniques for nucleic acid amplification and characterization combined with automation and user-friendly software have significantly broadened the diagnostic arsenal for the clinical microbiologist. The conventional diagnostic model for clinical microbiology has been labor-intensive and frequently required days to weeks before test results were available. Moreover, due to the complexity and length of such testing, this service was usually directed at the hospitalized patient population. The physical structure of laboratories, staffing patterns, workflow, and turnaround time all have been influenced profoundly by these technical advances. Such changes will undoubtedly continue and lead the field of diagnostic microbiology inevitably to a truly modern discipline. Advanced Techniques in Diagnostic Microbiology provides a comprehensive and up-to-date description of advanced methods that have evolved for the diagnosis of infectious diseases in the routine clinical microbiology laboratory. The book is divided into two sections. The first techniques section covers the principles and characteristics of techniques ranging from rapid antigen testing, to advanced antibody detection, to in vitro nucleic acid amplification techniques, and to nucleic acid microarray and mass spectrometry. Sufficient space is assigned to cover different nucleic acid amplification formats that are currently being used widely in the diagnostic microbiology field. Within each technique, examples are given regarding its application in the diagnostic field. Commercial product information, if available, is introduced with commentary in each chapter. If several test formats are available for a technique, objective comparisons are given to illustrate the contrasts of their advantages and disadvantages. The second applications section provides practical examples of application of these advanced techniques in several "hot" spots in the diagnostic field. A diverse team of authors presents authoritative and comprehensive information on sequence-based bacterial identification, blood and blood product screening, molecular diagnosis of sexually transmitted diseases, advances in mycobacterial diagnosis, novel and rapid emerging microorganism detection and genotyping, and future directions in the diagnostic microbiology field. We hope our readers like this technique-based approach and your feedback is highly appreciated. We want to thank the authors who devoted their time and efforts to produce their chapters. We also thank the staff at Springer Press, especially Melissa Ramondetta, who initiated the whole project. Finally, we greatly appreciate the constant encouragement of our family members through this long effort. Without their unwavering faith and full support, we would never have had the courage to commence this project.




The Evaluation of Forensic DNA Evidence


Book Description

In 1992 the National Research Council issued DNA Technology in Forensic Science, a book that documented the state of the art in this emerging field. Recently, this volume was brought to worldwide attention in the murder trial of celebrity O. J. Simpson. The Evaluation of Forensic DNA Evidence reports on developments in population genetics and statistics since the original volume was published. The committee comments on statements in the original book that proved controversial or that have been misapplied in the courts. This volume offers recommendations for handling DNA samples, performing calculations, and other aspects of using DNA as a forensic toolâ€"modifying some recommendations presented in the 1992 volume. The update addresses two major areas: Determination of DNA profiles. The committee considers how laboratory errors (particularly false matches) can arise, how errors might be reduced, and how to take into account the fact that the error rate can never be reduced to zero. Interpretation of a finding that the DNA profile of a suspect or victim matches the evidence DNA. The committee addresses controversies in population genetics, exploring the problems that arise from the mixture of groups and subgroups in the American population and how this substructure can be accounted for in calculating frequencies. This volume examines statistical issues in interpreting frequencies as probabilities, including adjustments when a suspect is found through a database search. The committee includes a detailed discussion of what its recommendations would mean in the courtroom, with numerous case citations. By resolving several remaining issues in the evaluation of this increasingly important area of forensic evidence, this technical update will be important to forensic scientists and population geneticistsâ€"and helpful to attorneys, judges, and others who need to understand DNA and the law. Anyone working in laboratories and in the courts or anyone studying this issue should own this book.




Diagnostic Molecular Biology


Book Description

Diagnostic Molecular Biology, Second Edition describes the fundamentals of molecular biology in a clear, concise manner with each technique explained within its conceptual framework and current applications of clinical laboratory techniques comprehensively covered. This targeted approach covers the principles of molecular biology, including basic knowledge of nucleic acids, proteins and chromosomes; the basic techniques and instrumentations commonly used in the field of molecular biology, including detailed procedures and explanations; and the applications of the principles and techniques currently employed in the clinical laboratory. Topics such as whole exome sequencing, whole genome sequencing, RNA-seq, and ChIP-seq round out the discussion. Fully updated, this new edition adds recent advances in the detection of respiratory virus infections in humans, like influenza, RSV, hAdV, hRV but also corona. This book expands the discussion on NGS application and its role in future precision medicine. - Provides explanations on how techniques are used to diagnosis at the molecular level - Explains how to use information technology to communicate and assess results in the lab - Enhances our understanding of fundamental molecular biology and places techniques in context - Places protocols into context with practical applications - Includes extra chapters on respiratory viruses (Corona)